1270576 五、發明說明（1) 【發明所屬之技術領域】 本發明係有關一種細胞培養裝置及方法，係可用於培 養任何微生物及細胞，以及收集其所生成之任何副產物。 【先前技術】 大規模的細胞培養程序已經於近年來被廣泛的開發並 使用於培養細菌、酵母菌與黴菌等擁有強韌的細胞壁及/ 或細胞外間質（e X t r a c e 1 1 u 1 a r m a t e r i a 1 )、較具彈性 的細胞。微生物細胞的彈性結構係此類高效率細胞培養程 序能夠快速發展的關鍵因素，例如：細菌細胞可於大量的 培養基中利用激烈的震動、搖晃以及通氣技術使其充分接 觸空氣，並維持良好的生長情形。另外，細菌也可於生物 膜上生長，不過需要一提供生長的平面。 相反地，培養真核細胞、動物細胞、哺乳動物細胞及 /或組織的技術就困難而複雜的多，原因在於此類細胞較 微生物細胞來的脆弱，並且其生長所需之養分與氧氣之維 持也較為困難而複雜。另外，跟微生物細胞不同的是，動 物細胞及/或哺乳動物細胞不能忍受通入空氣或混合性氣 體，例如：一混合性氣體包含氧氣、氮氣與二氧化碳，所 造成的劇烈的擾動及/或剪應力（s h e a r i n g f 〇 r c e )。並 且，動物細胞不能直接接觸空氣，多數動物細胞僅能利用 溶解於培養基中的氧氣。比起微生物細胞，動物細胞及哺 乳動物細胞更容易被氣體或是於通氣過程中遭受傷害，因 此增加其死亡率。應用於大規模培養之生物反應器通常具 備一個内部運轉器，例如：攪拌翼（i mpe 1 1 er )，使細胞1270576 V. INSTRUCTION DESCRIPTION OF THE INVENTION (1) Technical Field The present invention relates to a cell culture apparatus and method which can be used for cultivating any microorganisms and cells, and collecting any by-products thereof. [Prior Art] Large-scale cell culture procedures have been extensively developed in recent years and used to culture strong cell walls and/or extracellular stroma of bacteria, yeasts, and molds (e X trace 1 1 u 1 armateria) 1) More flexible cells. The elastic structure of microbial cells is a key factor in the rapid development of such high-efficiency cell culture procedures. For example, bacterial cells can be exposed to air and maintain good growth in a large amount of medium using intense vibration, shaking and aeration techniques. situation. In addition, bacteria can also grow on biofilms, but require a plane that provides growth. Conversely, techniques for culturing eukaryotic cells, animal cells, mammalian cells, and/or tissues are difficult and complex because of the fragility of such cells compared to microbial cells and the maintenance of nutrients and oxygen required for their growth. It is also difficult and complicated. In addition, unlike microbial cells, animal cells and/or mammalian cells cannot tolerate the introduction of air or mixed gases, for example, a mixed gas containing oxygen, nitrogen and carbon dioxide, causing severe disturbances and/or shears. Stress (shearingf 〇rce ). Moreover, animal cells are not exposed to air directly, and most animal cells can only utilize oxygen dissolved in the medium. Animal cells and mammalian cells are more susceptible to damage from gases or during aeration than microbial cells, thereby increasing their mortality. Bioreactors for large-scale cultures usually have an internal actuator, such as a stirring wing (i mpe 1 1 er ) to make cells
第5頁 1270576 五、發明說明（2) 承受非常高的液體剪應力而造成細胞的傷害或死亡，導致 細胞的低存活率及其蛋白質及/或副產品之產量也隨之降 低。同樣的，為使細胞均勻懸浮及/或適當接觸空氣，而 利用他種機械裝置、激烈的氣體運動或強烈的液體運動之 生物反應器也將造成細胞的傷害與妨礙細胞或組織之生 長，進一步造成細胞的副產品，例如：蛋白質，產量之減 少 〇 生物反應器主要之功能在於配合研究需要培養大量細 胞以萃取其衍生之微量活性物質，包括但不限於，細胞分 泌到培養基中的蛋白質或抗體；生物反應器之另一功能在 於大規模的細胞培養用以量產細胞或組織生產之活性蛋白 質以配合商業上的需求。由於真核細胞及/或原核細胞及/ 或動物細胞及/或哺乳動物細胞於實驗室大量培養的需 要，生物反應器及培養裝置在研究與生產細胞以製造活性 蛋白質及/或抗體及/或任何細胞副產品的方面扮演了重要 的角色。 許多習知方法可用來大規模或小規模培養細胞。在小 規模培養方面，近年來已發展出許多培養的容器，例如： 培養孤即是一例。培養皿基本上包含一可用來盛裝培養基 之底盤，以及一可掀開之上蓋。雖然可掀開之上蓋的確方 便操作，然而在培養過程中頻繁的掀蓋動作卻也常使細胞 遭到微生物的污染。而事實上，如何避免污染是一成功的 細胞或組織培養技術所面臨之最大挑戰。 為避免使用培養皿常發生的污染問題，於是發展出培Page 5 1270576 V. INSTRUCTIONS (2) The damage or death of cells caused by very high liquid shear stress causes the low cell survival rate and the production of protein and/or by-products to decrease. Similarly, in order to evenly suspend cells and/or properly contact the air, bioreactors that use their mechanical devices, intense gas movements or strong liquid motion will also cause cell damage and hinder the growth of cells or tissues, further The main function of the bioreactor is to produce a by-product of the cell, such as protein. The main function of the bioreactor is to cultivate a large number of cells to extract the trace active substances derived therefrom, including but not limited to proteins or antibodies secreted by the cells into the medium; Another function of bioreactors is in large-scale cell culture to produce active proteins produced by cells or tissues to meet commercial needs. Due to the need for large-scale culture of eukaryotic cells and/or prokaryotic cells and/or animal cells and/or mammalian cells in a laboratory, bioreactors and culture devices are researching and producing cells to produce active proteins and/or antibodies and/or Any aspect of cell byproducts plays an important role. Many conventional methods are available for culturing cells on a large scale or on a small scale. In terms of small-scale culture, many cultured containers have been developed in recent years, for example, cultivation is an example. The culture dish basically comprises a chassis for holding the culture medium, and a top cover can be opened. Although the upper cover can be opened easily, the frequent capping during the cultivation process often causes the cells to be contaminated by microorganisms. In fact, how to avoid contamination is the biggest challenge for a successful cell or tissue culture technology. In order to avoid the pollution problems that often occur in the culture dish, the development of cultivation
第6頁 1270576 五、發明說明（3) 養瓶（culture flasks)。培養瓶基本上含有一個培養 室、一管狀開口位於瓶子的一端及一對應之蓋子。此種設 計係為了減少細胞接觸到灰塵、細菌、酵母菌及其他污染 源的機會。教導培養瓶的專利係有，例如：C a r v e r公司所 擁有的美國專利第4, 334, 028號、Akamine公司所擁有的美 國專利第4, 851，351號及Degrassi所擁有的美國專利第 5，3 9 8，8 3 7號。雖然培養瓶較培養皿來得進步，其仍然無 法完全解決污染的問題。此外，培養孤與培養瓶均無法提 供充足的空氣，並且培養瓶所提供的生長面積並不如培養 皿來的充足，因此限制了培養的規模。 另一種用來培養細胞與組織之技術係為滾動式培養瓶 (r ο 1 1 e r b 〇 11 1 e s )，廣泛利用滾動式培養瓶技術於細胞 培養已有數年，雖然它具有某些培養皿或培養瓶所沒有之 優點，例如：提供細胞較大的生長與附著面積，然而還是 無法克服所有的缺點尤其是關於擴大培養規模方面。總而 言之，這些缺點包括，但不限制於，因為頂部空間之氣體 (gas headspace)與大量旋轉搜拌所造成的巨大流體剪 應力。由於滾動式培養瓶的高剪應力環境，導致較大的三 維結構的組織培養近乎不可能。唯有可以不受剪應力的傷 害及/或能維持貼附於滾動式培養瓶内壁的細胞才得以持 續培養一段時間。由此可知，維持長時間於滾動式培養瓶 培養一細胞株的困難點在於其高剪應力環境與污染的可能 性。關於滾動式培養瓶的例子包括：M u s s i公司所擁有的 美國專利第5, 5 2 7, 7 0 5號及Serkes公司所擁有的美國專利Page 6 1270576 V. Description of invention (3) Culture flasks. The culture bottle basically contains a culture chamber, a tubular opening at one end of the bottle and a corresponding lid. This design is designed to reduce the chance of cells coming into contact with dust, bacteria, yeast and other sources of pollution. The patents that teach the culture of the bottle are, for example, U.S. Patent No. 4,334,028 to the company of the company, U.S. Patent No. 4,851,351 to Akamine, and U.S. Patent No. 5 to the Degrassi. 3 9 8,8 3 No. 7. Although the culture bottle is more advanced than the culture dish, it still cannot completely solve the pollution problem. In addition, the culture of the solitary and the culture flasks does not provide sufficient air, and the growth area provided by the culture bottles is not as sufficient as that of the culture dishes, thus limiting the scale of the culture. Another technique used to culture cells and tissues is the rolling culture flask (r ο 1 1 erb 〇11 1 es ), which has been widely used in cell culture for several years, although it has some culture dishes or The advantages that the culture bottle does not have, for example, provide a large growth and attachment area of the cells, but it is still impossible to overcome all the disadvantages, especially regarding the expansion of the culture scale. In summary, these disadvantages include, but are not limited to, the large fluid shear stress caused by the gas headspace and the large number of rotating feeds. Due to the high shear stress environment of the rolling culture flask, tissue culture of larger three-dimensional structures is nearly impossible. Only cells that are not subject to shear stress and/or that remain attached to the inner wall of the roller culture bottle can be cultured for a period of time. From this, it is understood that the difficulty in maintaining a cell strain for a long time in a rolling culture flask lies in its high shear stress environment and the possibility of contamination. Examples of rolling culture flasks include: U.S. Patent No. 5, 5 2, 7 0 5 owned by Mus s i and US patents owned by Serkes
第7頁 1270576 五、發明說明（4) 第 4，9 6 2，0 3 3 號。 此外，雖然滾動 養瓶，然而其所提供 培養皿或培養瓶所提 故此，針對提高滾動 如，S e r k e s公司所擁 一種可增進細胞附著 式培養瓶的内壁加上 瓶之内表面積以供細 提供約850-1700平方 培養瓶才足以達到量 之培養系統雖然可節 貴及具限制性。 除了流體剪應力 與保持充分的氧氣供 題。正如本技術領域 胞，包括動物細胞、 黴菌的生長均有一主 step )，即氧質量轉 除了某些進行發 菌，以外，對於多數 對於細胞的代謝功能 細胞與動物細胞之培 裂的早期階段又特別 式培養瓶的整體面積大於培養皿或培 給每個細胞附著的面積並不一定大於 供之面積，特別是在大規模培養時。 培養瓶所能提供的生長面積的發明例 有的美國專利第5，0 1 0，0 1 3號係描述 表面積的滾動式培養瓶，其係於滾動 一種縐折狀的通道，增加滾動式培養 胞附著。然而，一般滾動式培養瓶僅 公分之面積，故仍需要大量的滾動式 產。而自動化、大量的滾動式培養瓶 省時間與人力，然而其操作卻相當昂 的問題與生 應也是細胞 之人士所熟 哺乳動物細 要的速率限 換(oxygen 酵作用的真 的原核細胞 是相當重要 養技術，氧 的重要。當 長面積 與組織 知，原 胞、昆 制步驟 mass 核微生 與真核 的，尤 氣的供 細胞懸 的限制之外 培養技術的 核細胞、真 蟲細胞、酵 ’獲得 中心問 核細 母菌與 rate-1i m i t i ng transfer ) 物，例如： 細胞，氧氣 其是針對哺 應對於細胞 浮時，對氧 酵母 的代謝 乳動物 快速分 氣的需 _画Page 7 1270576 V. Description of invention (4) No. 4, 9 6 2, 0 3 3. In addition, although the rolling bottle, however, is provided by the culture dish or the culture bottle, for the purpose of improving the rolling, for example, the internal wall of the cell-adjustable culture bottle which Serkes has can be added to the inner surface area of the bottle for fine provision. A culture system that is sufficient to achieve a quantity of about 850-1700 square flasks is expensive and restrictive. In addition to fluid shear stress and maintaining adequate oxygen supply. As in the field of technology, including the growth of animal cells and molds, there is a main step), that is, the oxygen mass is removed from some of the bacteria, and in the early stages of most of the cell-metabolic cell and animal cells. The overall area of the special culture flask is larger than the area in which the culture dish or culture is attached to each cell and is not necessarily larger than the area supplied, especially in large-scale culture. Inventive example of a growth area that can be provided by a culture bottle. U.S. Patent No. 5,010,013 is a rolling culture bottle for describing a surface area, which is rolled in a meandering passage to increase rolling culture. Cell attachment. However, in general, rolling flasks are only a few centimeters in area, so a large amount of rolling production is still required. Automated, large-scale rolling culture bottles save time and manpower. However, the operation is quite a problem. The rate limit is the same as that of the mammals who are familiar with the cells. (The true prokaryotic cells of the oxygen fermentation are quite Important nutrient technology, the importance of oxygen. When the long area and the tissue are known, the protocells, the Kuns step, the mass nuclear micronucleus and the eukaryotic, the nucleus of the cells, the culture of the nuclear cells, the true worm cells, Fermentation 'acquisition of the center of the nucleus and rate-1i miti ng transfer), such as: cells, oxygen is the need to feed the cell to float, the metabolism of milk yeast to the oxygen yeast quickly
-n 第8頁 1270576 五、發明說明（5) 求量最大，、而當細胞聚集或是分化後，氧需求量就會降 低。有些哺乳動物細胞與動物細胞為附著依存 (anchorage-dependent )，另外有些哺乳動物細胞與動 物細胞則可於液體環境中懸浮生長，即，非附著依存 (anchorage independent )，然而所有的細胞均需生長 於充分的溶氧環境甲。在細胞培養的後期，不論是附著依 存或是非附著依存的細胞，每單位體積的細胞數量會大量 增加，因而再度需要大量的氧質量轉換以提供足夠&氣。 基本上，至少對於附著生長的細胞，可以利用機器攪 拌以及通入氣體的方式來供應其需求的氧氣，然而，如前 所述，攪拌和通氣的動作均會傷害細胞，因而減低其存活 率及整體細胞及/或組織培養的的效率與產量。此外，直 接通入氣體到細胞及組織培養基中會導致氣泡產生，同樣 不利於細胞存活。 為解決細胞培養時氧氣提供的問題而發明的裝置，例 如：W ο 1 f e t a 1 ·所擁有的美國專利第5，1 5 3，1 3 1號，係關 於一種沒有攪拌翼（blades)的生物反應槽（bioreactor vessel )。此生物反應槽係將空氣由輸氣通道（a i r inlet passageway )通入，經由一支持板元件(support plate member)穿過屏幕（screen)，並通過夾於反應槽 外箱（h 〇 u s i n g )兩側間的一扁盤狀之滲透膜（f 1 a t d i s k permeable membrane )，利用外箱兩側之間的氧濃度梯 度，使氧氣可以擴散通過滲透膜而到達培養室（culture chamber ) 〇-n Page 8 1270576 V. INSTRUCTIONS (5) The largest amount is obtained, and when cells are aggregated or differentiated, the oxygen demand is reduced. Some mammalian cells are anchorage-dependent with animal cells. In addition, some mammalian cells and animal cells can be suspended in a liquid environment, that is, anchorage independent, but all cells need to grow. In a full dissolved oxygen environment A. In the later stages of cell culture, the number of cells per unit volume is greatly increased, whether it is adherent or non-adherent-dependent cells, and thus a large amount of oxygen mass conversion is required again to provide sufficient & gas. Basically, at least for adherently growing cells, the required oxygen can be supplied by means of machine agitation and gas injection. However, as mentioned above, both agitation and aeration can damage the cells, thereby reducing their survival rate and Efficiency and yield of whole cell and/or tissue culture. In addition, direct gas flow into cells and tissue culture media can cause bubbles to form, which is also detrimental to cell survival. A device invented to solve the problem of oxygen supply during cell culture, for example: W ο 1 feta 1 · U.S. Patent No. 5,1 5 3,1 31, which is related to a creature without a blade. Reaction vessel (bioreactor vessel). The bioreactor channel passes air through an air inlet passageway, passes through a screen via a support plate member, and passes through a reaction box (h 〇using ) A flat disk-shaped permeable membrane between the sides, using the oxygen concentration gradient between the two sides of the outer tank, allows oxygen to diffuse through the permeable membrane to reach the culture chamber.
第9頁 1270576 五、發明說明（6) 然而，Wo 1 f的生物反應槽具有許多缺點，尤其是，氧 氣經由扁盤狀之滲透膜擴散的速率成為培養室.規模的主要 限制因素，另外一個缺點是扁盤狀之滲透膜設計成可伸縮 以具有攪拌功能，此點卻可能造成細胞的死亡。利用攪拌 功能以使空氣均勻分布於培養基中是相當重要的，然而， 攪拌的同時也將增加培養室内部的剪應力，如前所述可能 造成對細胞的傷害。因此，當設計生物反應器或培養槽 時，如何提供足夠的氣體交換以維持培養較大的細胞結 構，係相當重要並且為一實際的限制。 為了解決前述缺點的裝置，利用透氣材料（g a s permeable material )戶斤製造的反應器例如：Schwarz e t a 1.所擁有的美國專利第5，7 0 2，9 4 1號，名稱為π可透氣的 生物反應器及其使用方法"，係關於一種可水平轉動的容 器，並且其容器壁係部分採用透氣材料，以期達到直接透 過此透氣材料與培養基進行氣體交換之目的。 然而，Schwarz所揭露之裝置的規模仍然受到限制， 原因在於氣體交換需依據透氣面積的大小來決定，Page 9 1270576 V. INSTRUCTIONS (6) However, the biological reaction tank of Wo 1 f has many disadvantages. In particular, the rate of diffusion of oxygen through the flat disk-shaped permeable membrane becomes the main limiting factor of the culture chamber. The disadvantage is that the flat disk-shaped permeable membrane is designed to be stretchable to have a stirring function, which may cause cell death. It is important to use the agitation function to evenly distribute the air in the medium. However, the agitation will also increase the shear stress inside the chamber, which may cause damage to the cells as described above. Therefore, when designing a bioreactor or culture tank, how to provide sufficient gas exchange to maintain a larger cell structure is important and a practical limitation. In order to solve the aforementioned disadvantages, a reactor made of a gas permeable material, for example, Schwarz eta 1. U.S. Patent No. 5,7 0 2,9 4 1 is known as π breathable. The bioreactor and method of use thereof relates to a horizontally rotatable container, and the wall portion of the container is made of a gas permeable material, so as to achieve gas exchange directly with the medium through the gas permeable material. However, the size of the device disclosed by Schwarz is still limited because gas exchange is determined by the size of the gas permeable area.
Schwarz雖然強調當裝置的表面積增加，培養基的體積量 亦相對增加，而於其說明書中所揭露之較佳實施例之規模 限制於直徑1至6英吋，寬度則介於1 / 4至1英吋間，這樣的 大小並不合適生長具有三維空間的細胞集結物與組織及/ 或任何大量規模生產。 同樣地，F a 1 k e n b e r g e t a 1.所擁有的美國專利第 5，4 4 9，6 1 7號，標題為n用於細胞培養之培養容器n ，係關Although Schwarz emphasizes that the surface area of the device increases as the surface area of the device increases, the size of the preferred embodiment disclosed in the specification is limited to a diameter of 1 to 6 inches and a width of between 1 / 4 and 1 inch. In the meantime, such a size is not suitable for growing cell aggregates and tissues with three-dimensional space and/or any large-scale production. Similarly, F a 1 k e n b e r g e t a 1. The U.S. Patent No. 5,4 4 9,6,7, entitled, n, is used for cell culture culture vessels n,
1270576 五、發明說明（7) 於一水平旋轉之裝置，此培養容器藉由一透析膜將内部區 分為細胞培養腔室與營養液儲存槽（nutrient medium reservoir )，透氣材料用在本容器壁使得細胞培養室可 進行氣體交換，不過營養液儲存槽也並非完全充滿營養 液，而是兩個腔室的培養液上方都維持大量的氣體。然 而，F a 1 k e n b e r g容器並未設計用以減少細胞培養室内部的 擾動，相反的，為了保持透析膜的潮濕，攪拌是必要的步 驟；此外，F a 1 k e n b e r g並未提及使用此容器於培養細胞集 結物或是任何類型的組織。 為解決氧氣供應的問題之發明還有，例如：L i au所擁 有的美國專利第5，7 6 6，9 4 9號，標題為"培養附著依存之單 層細胞的方法與裝置π ，係描述一種上下震盪培養基以增 加氧氣提供的培養系統。 然而L i a u的發明具有很多缺點，其中之一就是其裝置 的複雜性。L i a u的系統包括兩個外接的儲存槽以及一個含 有一系歹1J 垂直基材平盤(vertical substrate plates ) 的分離式培養室，另外還需要數個蠕動式幫浦 (peristaltic pumps )推動培養液由一個儲存槽流經培 養室再流到另一個儲存槽，最後再流回到第一個儲存槽。 因為L i a u的裝置複雜而且有一部份為培養室外部的設備， 例如··外部管線、儲存槽與幫浦，所以極有可能將外界的 污染源導入内部，另外，因為裝置的設備龐大，滅菌工作 不易進行又耗費人力。 L i a u的發明另一個缺點是培養液流經系統時所造成的1270576 V. INSTRUCTION DESCRIPTION (7) In a horizontally rotating device, the culture vessel is internally divided into a cell culture chamber and a nutrient medium reservoir by a dialysis membrane, and a gas permeable material is used on the wall of the container. The cell culture chamber can exchange gas, but the nutrient solution storage tank is not completely filled with nutrient solution, but a large amount of gas is maintained above the culture liquid of the two chambers. However, the F a 1 kenberg container is not designed to reduce disturbances inside the cell culture chamber. Conversely, agitation is a necessary step in order to keep the dialysis membrane moist; in addition, F a 1 kenberg does not mention the use of this container. Culture cell aggregates or any type of tissue. In order to solve the problem of the supply of oxygen, for example, U.S. Patent No. 5,7,6,9,9,9, entitled, "Method and Apparatus for Cultivating Attached Dependent Monolayer Cells, A culture system in which an up and down shaking medium is added to increase oxygen supply is described. However, the invention of L i a u has many disadvantages, one of which is the complexity of its device. L iau's system consists of two external storage tanks and a separate culture chamber containing a series of 1J vertical substrate plates. In addition, several peristaltic pumps are required to drive the culture solution. A storage tank flows through the culture chamber and then flows to another storage tank, and finally flows back to the first storage tank. Because the device of Liau is complicated and there is a part of the equipment outside the culture, such as external pipelines, storage tanks and pumps, it is very possible to introduce external pollution sources into the interior. In addition, because the equipment of the device is huge, sterilization work It is difficult to carry out and labor intensive. Another disadvantage of the invention of L i a u is caused by the flow of culture fluid through the system.
第11頁 1270576 五、發明說明（8) 流體剪應力 胞，因而減 並不利於細 並堆積在平 低細胞的存 新開始，相 複雜性，收 耗時。最後 將會暴露於 細胞死亡。 研究、患者 技術之缺陷 揭露一種細 不複雜、更 加細胞存活 以滿足長久 【發明内容 本發明 其產物之可 即丟與有效 可能會 低細胞 胞附著 盤的底 活率以 當沒有 集任何 ，當生 空氣中 細胞與 護理與 與限制 胞和組 有效率 率與細 以來的 干擾 的存 ，因 端， 及蛋 效率 分泌 長液 ， 即 組織 學術 並且 織培 、較 胞副 需要 並且 活率 為某 這些 白質 而且 的蛋 低於 ，直 培養 研究 補足 養之 省事 產物 移動生 。另外 些無法 細胞最 產量， 不理想 白質或 細胞生 接接觸 技術對 都相當 其不便 方法與 、較不 的產量 長於 ，垂 即刻 後大 而此 〇再 是細 長的 氣體 於生 的重 之處 裝置 曰 主 叩貝 之一 基材 直基 附著 多會 系統 者， 胞產 基質 環境 物科 要。 ，本 ，提 、較 種新 平盤 材平 的細 死亡 必須 由於 物都 平盤 ，因 技研 為了 發明 供更 省人 方法 上的 盤的 胞會 ，因 一再 此系 很麻 時， 此將 究、 克服 係教 可靠 力並 與裝 細 設計 掉落 此降 地重 統的 煩且 細胞 導致 藥物 習知 導並 、較 可增 置， 係提供一種用來 靠、簡易、便宜 率的方法與裝置 與揭露一種新穎的方法與裝 原核細胞、真核 胞、細菌或黴菌 細胞直接接觸空 胞，例如： 胞、人類細 不會讓任何 培養細胞及/或組織以及收集 、可循環使用及/或可以用完 。更具體而言，本發明係教導 置可以有效的培養欲培養之細 細胞、動物細胞、哺乳動物細 ，可以持續提供氧氣和養分卻 氣，因此可以降低細胞的傷害Page 11 1270576 V. INSTRUCTIONS (8) Fluid shear stress cells, which are not conducive to thinning and accumulation at the beginning of the development of flat cells, complexity, and consumption. Eventually it will be exposed to cell death. The defects of research and patient technology reveal a kind of fine and uncomplicated, more cell survival to satisfy the long-term [invention content, the product of the present invention can be thrown away and effectively can lower the cell bottom of the cell to adhere to the disk when there is no set, when the In the air, cells and care and limiting the efficiency of cells and groups, the interference of the end, the end, and the efficiency of the egg secretion, that is, the organization of academic and woven, more than the side needs and the rate of some of these white matter In addition, the egg is lower than the direct cultivation research to make up for the survival of the product. Others are unable to produce the most cells, and the undesired white matter or cell-to-contact technology is quite inconvenient and less prolific. The yield is longer than that of the sputum. One of the main mussels is a substrate with a direct attachment to the multi-system. , Ben, mention, and the new type of flat plate, the fine death of the flat must be due to the flatness of the goods, because the technical research in order to invent the disk for the more provincial methods, because this line is very numb, this will be studied, overcome Teach the reliability and the fine design to drop the nuisance of the descending and the cells lead to the drug's conventional guidance, and can be added, providing a method and device for relying on, simple and cheap, and exposing a kind The novel method directly contacts the empty cells with prokaryotic, eukaryotic, bacterial or fungal cells, for example: cells, human fines do not allow any cultured cells and/or tissues to be collected, recyclable, and/or run out. More specifically, the present invention teaches that the cells, animal cells, and mammals to be cultured can be effectively cultured, and oxygen and nutrients can be continuously supplied, thereby reducing cell damage.
第12頁 1270576 五、發明說明（9) 甚至細胞死亡率。此外，本發明之方法與裝置係可減少污 染，避免細胞於提供氣體時直接受到剪應力的傷害，因而 可以預防氣泡與氣體對細胞的負面影響。再者，本發明之 裝置係提供一更簡單、更便利之方法生產與收集細胞產 物，例如：蛋白質，及/或抗體，及/或病毒，及/或任何 細胞及/或組織之產物。同時，藉由氣體出口壓力感測與 調控設備以便造成培養腔室内壓力之固定或梯度變化，利 用此變化將有利於特定細胞之生長與分化。 本發明之目的係包括，但不限於：提供一新穎之方法 與裝置以製備、生長與維持細胞；提供一新穎之方法與裝 置以生長及/或製備組織培養物；提供一新穎之方法與裝 · 置以製備及生長器官培養物；提供一新穎之方法與裝置， 在充足的氧氣與養分供應及最少的污染之環境下培養細 胞；提供一新穎之方法與裝置在一多孔的生長基材 (porous growth substrate )上培養細胞，以力口強細胞 的氧氣與養分之交換，卻不會因為注入氣體而使細胞直接 接觸到氣泡及/或剪應力；提供一生長基材以強化細胞的 附著力、增加空氣與培養液的接觸面積，以及作為一個裝 置中靜態的培養基攪拌器；提供一方法與裝置可以使細胞 污染減至最低，提供一方法與裝置可使細胞死亡率減至最 低，因而增加細胞產物之產量，提供一方法與裝置可以使 φ 培養時的流體剪應力大量減少甚至完全沒有，使細胞死亡 率減至最低；提供一方法與裝置在培養時不需要藉由通氣 泡來供應培養基氧氣；提供一方法與裝置使培養基可以移Page 12 1270576 V. Description of invention (9) Even cell death rate. In addition, the method and apparatus of the present invention can reduce contamination and prevent cells from being directly subjected to shear stress when supplying gas, thereby preventing the negative effects of bubbles and gases on the cells. Furthermore, the device of the present invention provides a simpler and more convenient means of producing and collecting cellular products, such as proteins, and/or antibodies, and/or viruses, and/or products of any cells and/or tissues. At the same time, the use of gas outlet pressure sensing and conditioning equipment to cause a fixed or gradient change in the pressure within the culture chamber will facilitate the growth and differentiation of specific cells. The present invention includes, but is not limited to, providing a novel method and apparatus for preparing, growing and maintaining cells; providing a novel method and apparatus for growing and/or preparing tissue culture; providing a novel method and apparatus · Preparing and growing organ cultures; providing a novel method and apparatus for culturing cells in an environment of sufficient oxygen and nutrient supply with minimal contamination; providing a novel method and apparatus in a porous growth substrate The cells are cultured on a porous growth substrate, and the oxygen and nutrients of the cells are strongly exchanged, but the cells are not directly exposed to bubbles and/or shear stress by injecting gas; a growth substrate is provided to enhance cell adhesion. Force, increasing the contact area of air with the culture fluid, and acting as a static medium agitator in a device; providing a method and apparatus to minimize cell contamination, providing a means and means to minimize cell death, thus Increasing the yield of cell products, providing a method and apparatus for greatly reducing the fluid shear stress during φ culture To no, the cell death rate to a minimum; providing a method and apparatus need not be supplied by aeration bubbles in the culture medium of oxygen; to provide a method and apparatus so that the medium can move
第13頁 1270576 五、發明說明（ίο) 動，讓掉落的細胞可以再次分配並且幫助增進細胞的附著 能力；提供一方法與裝置可持續供應細胞培養基與氧氣， 使細胞產物之產量達到最大量；提供更多的面積讓細胞附 著；提供一方法與裝置可有效率的製備與收集細胞及/或 組織分泌的蛋白質與其他產物；以及提供一培養細胞之方 法與裝置，使得培養一段時間後細胞的生存率可獲得改 善。提供一種方法與裝置可使培養腔室内的壓力可以隨細 胞之特性加以調整，以促使細胞生長或/與分化。 本發明之主要目的係提供一種細胞培養裝置，其至少 包含：一個以上之腔室；一充填結構，用以充填生長基材 以接受至少一個以上之細胞以及允許細胞貼附及/或附 著；以及一氣泡排除結構，用以防止培養過程中產生之氣 泡干擾細胞及/或生長基材。 前述細胞培養裝置，至少包含：一内腔室；一外腔 室；一充填結構，用以填充生長基材以接受至少一個以上 之細胞以及允許細胞貼附及/或附著；以及一氣泡排除結 構，用以防止培養過程中產生之氣泡干擾細胞及/或生長 基材；其中前述内腔室與外腔室之間係可相互連通。 前述之充填結構之設置位置並不受限制，較佳係設置 於外腔室，且其填充之生長基材種類不受限制，可自由選 擇適合該培養細胞或組織之基材。 前述氣泡排除結構係包含：階梯式斜板結構、鰭片式 結構、排氣通道、壓力式感應閥件以及任何可排除氣泡產 生之類似結構單獨或搭配使用，其可選擇性地設置於内腔Page 13 1270576 V. Description of the invention (ίο), allowing the dropped cells to be redistributed and helping to improve the adhesion of the cells; providing a method and apparatus for the sustainable supply of cell culture medium and oxygen to maximize the production of cellular products Providing more area for cell attachment; providing a method and apparatus for efficiently preparing and collecting proteins and other products secreted by cells and/or tissues; and providing a method and apparatus for culturing cells such that cells are cultured for a period of time The survival rate can be improved. A method and apparatus are provided to allow the pressure within the culture chamber to be adjusted to the characteristics of the cells to promote cell growth or/and differentiation. A primary object of the present invention is to provide a cell culture apparatus comprising at least: one or more chambers; a filling structure for filling a growth substrate to receive at least one or more cells and allowing cells to attach and/or adhere; A bubble exclusion structure is used to prevent bubbles generated during the culture from interfering with cells and/or growing the substrate. The cell culture device comprises at least: an inner chamber; an outer chamber; a filling structure for filling the growth substrate to receive at least one of the cells and allowing the cells to attach and/or adhere; and a bubble exclusion structure The air bubble generated during the culture process is prevented from interfering with the cells and/or the growth substrate; wherein the inner chamber and the outer chamber are in communication with each other. The above-mentioned filling structure is not limited, but is preferably disposed in the outer chamber, and the type of the growth substrate to be filled is not limited, and the substrate suitable for the cultured cells or tissues can be freely selected. The bubble exclusion structure comprises: a stepped swash plate structure, a fin structure, an exhaust passage, a pressure sensing valve member, and any similar structure capable of eliminating air bubbles, which may be separately or in combination, which may be selectively disposed in the inner cavity.
第14頁 1270576 五、發明說明（11) 室或外腔室。 本發明之細胞培養裝置，進一步可包含：空氣幫浦、 進料幫浦、出料幫浦、培養基容器、產物收集容器、壓力 感應式閥件、計時器等單元。 藉由本發明之細胞培養裝置，可使培養基於腔室間來 回往返，以溫和地再次分配掉落的細胞回到生長基材上， 因而能強化細胞附著能力，可藉由混勻培養基而幫助氣體 與培養基的接觸，以及在不傷害或殺死細胞的前提下，確 保養分與氣體平均分布。Page 14 1270576 V. Description of the invention (11) Room or external chamber. The cell culture apparatus of the present invention may further comprise: an air pump, a feed pump, a discharge pump, a medium container, a product collection container, a pressure sensing valve member, a timer, and the like. By the cell culture device of the present invention, the medium can be reciprocated back and forth between the chambers to gently re-distribute the dropped cells back to the growth substrate, thereby enhancing the cell adhesion ability, and the gas can be assisted by mixing the medium. Ensure contact with the medium and ensure an even distribution of nutrients and gases without harming or killing the cells.
本發明之第一種實施態樣係為一細胞培養裝置，其至 少包含：一内腔室；一外腔室；一生長基材填充結構，係 設置於前述外腔室中；一階梯式斜板結構，係設置於前述 外腔室中，以阻隔培養過程中產生之氣泡；前述内腔室與 外腔室之間係可相互連通。 本發明之第二種實施態樣係為一種細胞培養裝置，其 至少包含：一内腔室；一外腔室；一生長基材填充結構， 係設置於前述外腔室中；一鰭片式結構，係設置於前述生 長基材填充結構壁緣，以阻隔培養過程中產生之氣泡；前 述内腔室與外腔室之間係可相互連通。A first embodiment of the present invention is a cell culture device comprising: at least: an inner chamber; an outer chamber; a growth substrate filling structure disposed in the outer chamber; a stepped oblique The plate structure is disposed in the outer chamber to block air bubbles generated during the cultivation process; the inner chamber and the outer chamber are connected to each other. A second embodiment of the present invention is a cell culture device comprising at least: an inner chamber; an outer chamber; a growth substrate filling structure disposed in the outer chamber; a fin type The structure is disposed on the edge of the growth substrate filling structure to block bubbles generated during the cultivation process; the inner chamber and the outer chamber are connected to each other.
本發明之第三種實施態樣係為一種細胞培養裝置，其 至少包含：一内腔室；一外腔室；一生長基材填充結構， 係設置於前述外腔室中；一排氣通道，係設置於前述内腔 室之頂部，以排除培養過程中產生之氣泡；前述内腔室與 外腔室之間係可相互連通。A third embodiment of the present invention is a cell culture device comprising at least: an inner chamber; an outer chamber; a growth substrate filling structure disposed in the outer chamber; and an exhaust passage The system is disposed at the top of the inner chamber to exclude air bubbles generated during the cultivation process; the inner chamber and the outer chamber are connected to each other.
第15頁 1270576 五、發明說明（12) 前述細胞培養裝置其進一步包含一浮板腸膜結構設置 於前述排氣通道下方，可將排氣通道阻斷，以防止培養基 流出内腔室而污染空氣濾網。 本發明之第四種實施態樣係為一種細胞培養裝置，其 至少包含：一内腔室；一外腔室；一生長基材填充結構’ 係設置於前述外腔室中；一壓力感應式閥件，係設置於前 述内腔室頂部，以阻隔培養過程中產生之氣泡；前述内腔 室與外腔室之間係可相互連通。 前述之壓力感應式閥件係可進一步搭配一計時器，使 該壓力感應式閥件採脈衝式啟閉。 前述各實施態樣可於空氣幫浦導入腔室過程t可以選 擇性的裝設一空氣濾網以防止污染。 本發明之細胞培養裝置係可作二維或三維組織之培 養。 本發明之腔室係可由任何材料構成，包括但不限於， 玻璃、金屬或高分子材質。 本發明使用之生長基材可為任何材質，包括但不限 於，例如：陶瓷、生物可分解基質、高分子聚合物、編織 基材(woven substrate )、非編織基材（non - woven substrate )、聚驢胺、聚酯、聚氨酯、碳氟聚合物、聚 乙烯、聚丙烯或聚乙烯醇、三甲胺、玻璃、矽與二乙氨乙 基（D E A E )。多孔生長基材可為任何型式、形狀或大小， 包括但不限於，碟狀、薄片、塊狀、盤狀、片狀、帶狀、 粒狀、半透性顆粒、半透性膜或半透性中空纖維。Page 15 1270576 V. The invention (12) The foregoing cell culture device further comprises a floating plate intestinal membrane structure disposed under the exhaust passage to block the exhaust passage to prevent the medium from flowing out of the inner chamber to pollute the air Filter. A fourth embodiment of the present invention is a cell culture device comprising: at least: an inner chamber; an outer chamber; a growth substrate filling structure disposed in the outer chamber; a pressure sensing type The valve member is disposed on the top of the inner chamber to block air bubbles generated during the cultivation process; the inner chamber and the outer chamber are connected to each other. The aforementioned pressure-sensing valve member can be further equipped with a timer to enable the pressure-sensing valve member to be pulsed to open and close. The foregoing embodiments can be used to selectively introduce an air filter to prevent contamination during the introduction of the air pump into the chamber. The cell culture apparatus of the present invention can be cultured in two or three-dimensional tissues. The chamber of the present invention may be constructed of any material including, but not limited to, glass, metal or polymeric materials. The growth substrate used in the present invention may be any material including, but not limited to, ceramics, biodegradable substrates, high molecular polymers, woven substrates, non-woven substrates, Polyamide, polyester, polyurethane, fluorocarbon polymer, polyethylene, polypropylene or polyvinyl alcohol, trimethylamine, glass, hydrazine and diethylaminoethyl (DEAE). The porous growth substrate can be of any type, shape or size, including but not limited to, dish, flake, block, disc, flake, ribbon, granule, semipermeable granule, semipermeable membrane or semipermeable Hollow fiber.
II 第16頁 1270576 五、發明說明（13) 本發明之另一目的係提供一種細胞培養方法，主要係 利用貼附及/或附著有細胞之生長基材，間歇性及/或定期 性的接觸空氣-培養基之界面以獲得氧氣，其特徵為藉由 一氣泡排除結構以避免培養過程產生之氣泡干擾細胞及生 長基材。 本發明係提供一種方法與裝置可使培養腔室内的壓力 可以隨所培養細胞之特性加以調整，以促使細胞生長或/ 與分化。 任何細胞均可生長於本發明之細胞培養裝置，例如： 原核細胞、真核細胞及/或哺乳動物細胞。此外，附著依 存或非附著依存之細胞也適用於本發明。附著依存細胞需 要表面附著生長，然而非附著依存細胞則生長於液狀懸浮 液中，所有種類的細胞均需要充足的氧氣、養分與生長因 子以供其生長。 本發明提供一新穎之方法，係可間歇性，但非直接 的，經由一空氣-培養基之薄層界面提供細胞氧氣，卻不 會傷害及/或殺死細胞。 【實施方式】 以下之實施例僅供說明而不宜用來侷限本發明之範 圍。發明之詳細說明可與所附之圖式結合參考，用以幫助 獲得更充分的了解。 實施例一 第一 A圖及第二B圖係顯示本發明之第一個實施例，一 種細胞培養裝置1 0 0，主要包含：一内腔室1 0 0 5 ; —外腔II Page 16 1270576 V. INSTRUCTION DESCRIPTION (13) Another object of the present invention is to provide a method of cell culture, mainly by using a growth substrate to which cells are attached and/or attached, intermittent and/or periodic contact. The air-medium interface is used to obtain oxygen, which is characterized by a bubble exclusion structure to prevent bubbles generated by the culture process from interfering with cells and growing the substrate. SUMMARY OF THE INVENTION The present invention provides a method and apparatus for adjusting the pressure within a culture chamber to the characteristics of the cultured cells to promote cell growth or/and differentiation. Any cell can be grown in the cell culture device of the present invention, for example, a prokaryotic cell, a eukaryotic cell, and/or a mammalian cell. Furthermore, adherent or non-adherent dependent cells are also suitable for use in the present invention. Attachment-dependent cells require surface attachment growth, whereas non-adherent cells grow in a liquid suspension, and all types of cells require sufficient oxygen, nutrients, and growth factors for growth. The present invention provides a novel method for intermittently, but not directly, providing cellular oxygen through a thin layer interface of air-medium without harming and/or killing cells. [Embodiment] The following examples are for illustrative purposes only and are not intended to limit the scope of the invention. Detailed descriptions of the invention can be combined with the accompanying drawings to help provide a better understanding. Embodiment 1 FIG. 1A and FIG. 2B show a first embodiment of the present invention, a cell culture device 100, which mainly comprises: an inner chamber 1 0 0 5;
第17頁 1270576 五、發明說明（14) 室1 0 0 4 ; —填充結構1 0 0 7，係設置於前述外腔室1 0 0 4中， 用以填充生長基材1 0 1 0 ; —階梯式斜板結構1 0 0 6，係設置 於前述外腔室1 0 0 4中，以阻隔培養過程中產生之氣泡；前 述内腔室1 0 0 5與外腔室1 0 0 4之間係可相互連通。此培養裝 置1 0 0進一步包含：感應式閥件1 0 0 1 、空氣幫浦1 0 1 1 、新 鮮培養基容器1 0 0 8、產物收集容器1 0 0 3、進料幫浦1 0 0 9、 出料幫浦1 0 0 2。本細胞培養裝置1 0 0進一步可於空氣幫浦 1 0 1 1導入腔室過程中可以選擇性的裝設一空氣濾網1 0 1 2以 防止污染。 此細胞培養裝置1 0 0之操作方式如下：如第一 A圖所 示，當空氣幫浦1 0 1 1未做動前，大部分之培養基位於下 層，因此生長基材1010暴露於空氣之中；如第一B圖所 示，當空氣幫浦1 0 1 1啟動後，内腔室1 0 0 5中之培養基會受 到空氣壓力，由内腔室1 0 0 5向外腔室1 0 0 4壓縮，促使培養 基上升於外腔室1 0 0 5而淹蓋住附著細胞之生長基材1 0 1 0 ; 當外腔室1 0 0 4之壓力值上升至一特定值之後，則關閉空氣 幫浦1 0 1 1並將壓力式感應閥件1 0 0 1打開，將外腔室1 0 0 4之 壓力釋放，並使外腔室1 0 0 4之培養基因重力流回内腔室 1 0 0 5中，此時生長基材1 0 1 0則又重新暴露於空氣中，如第 一A圖所示。如此反覆進行第一 A圖及第一 B圖之程序，係 可間歇性，但非直接的，經由一空氣-培養基之薄層界面 提供細胞氧氣，卻不會傷害及/或殺死細胞。同時，為避 免過量空氣吹入而導致培養基完全擠入外腔室1 〇 〇 4，並有 多餘空氣由内腔室1005推入外腔室1004而造成氣泡直接衝Page 17 1270576 V. Description of the invention (14) Room 1 0 0 4 ; - Filling structure 1 0 0 7 is placed in the aforementioned outer chamber 1 0 0 4 to fill the growth substrate 1 0 1 0; The stepped swash plate structure 100 is disposed in the outer chamber 1 0 0 4 to block bubbles generated during the cultivation process; between the inner chamber 1 0 0 5 and the outer chamber 1 0 0 4 The systems can be connected to each other. The culture device 100 further comprises: an inductive valve member 1 0 0 1 , an air pump 1 0 1 1 , a fresh medium container 1 0 0 8 , a product collection container 1 0 0 3, a feed pump 1 0 0 9 , discharge pump 1 0 0 2 . The cell culture device 100 can further selectively install an air filter 1 0 1 2 during the introduction of the air pump 1 0 1 1 to prevent contamination. The operation mode of the cell culture apparatus 100 is as follows: as shown in the first A diagram, most of the culture medium is located in the lower layer before the air pump 1 0 1 1 is not actuated, so the growth substrate 1010 is exposed to the air. As shown in the first B diagram, when the air pump 1 0 1 1 is started, the medium in the inner chamber 1 0 0 5 will be subjected to air pressure, and the inner chamber 1 0 0 5 is outwardly chamber 1 0 0 4 compression, causing the medium to rise in the outer chamber 1 0 0 5 and flooding the growth substrate of the attached cells 1 0 1 0; when the pressure value of the outer chamber 1 0 0 4 rises to a specific value, the air is turned off The pump 1 0 1 1 opens the pressure-sensing valve member 1 0 0 1 , releases the pressure of the outer chamber 1 0 0 4 , and causes the culture gene of the outer chamber 1 0 0 4 to flow back into the inner chamber 1 In 0 0 5, at this time, the growth substrate 1 0 10 is re-exposed to the air, as shown in FIG. The procedure of performing the first A map and the first B graph in this manner is intermittent, but not straightforward, providing cellular oxygen through a thin layer interface of air-medium without harming and/or killing the cells. At the same time, in order to avoid excessive air blowing, the medium is completely squeezed into the outer chamber 1 〇 〇 4, and excess air is pushed from the inner chamber 1005 into the outer chamber 1004, causing direct bubble rushing.
IIIIII
第18頁 1270576 五、發明說明（15) 擊生長基材1 0 1 0部分，因此利用一階梯式斜板結構1 0 0 6將 過剩氣泡導入外腔室1 0 0 4壁緣而阻絕氣泡直接衝擊細胞， 造成細胞損壞。 此外，利用進料幫浦1 0 0 9與出料幫浦1 0 0 2進行培養基 連續灌注培養。壓力感應式閥件1 0 0 1亦可永遠開啟，並以 計時器（圖未顯示）做為空氣幫浦1 0 1 1做動之依據。或 是，將計時器以及壓力感應式閥件1 0 0 1搭配使用，亦即壓 力感應閥件1 0 0 1採脈衝式啟閉，如此更可易於控制外腔室 1 0 0 4壓力，且當計時器時間到時亦可關閉空氣幫浦1 0 1 1 。 本發明之細胞培養裝置1 0 0係可選擇性的以自動化或以手 動方式操作。 實施例二 第二A圖及第二B圖係顯示本發明之第二個實施例，一 種細胞培養裝置2 0 0，其至少包含：一内腔室2 0 0 4 ; —外 腔室2 0 0 3 ; —填充結構2 0 0 5，係設置於前述外腔室2 0 0 3 中，用以填充生長基材2008 ; —向上斜之鰭片式結構 2 0 0 9，係設置於前述生長基材填充結構2 0 0 5壁緣，以阻隔 培養過程中產生之氣泡；前述内腔室2004與外腔室2003之 間係可相互連通。此培養裝置2 0 0進一步包含：感應式閥 件2 0 0 1、空氣幫浦2 0 0 7、新鮮培養基容器2 0 1 0、產物收集 容器2 0 1 1 、進料幫浦2 0 0 6、出料幫浦2 0 0 2。本細胞培養裝 置200進一步可於空氣幫浦2007導入腔室過程中可以選擇 性的裝設一空氣濾網2 0 1 2以防止污染。 此細胞培養裝置2 0 0之操作方式如下：如第二A圖所Page 18 1270576 V. Description of the invention (15) Strike the growth substrate 1 0 1 0 part, so use a stepped sloping plate structure 1 0 0 6 to introduce excess air bubbles into the outer chamber 1 0 0 4 wall edge to block the bubble directly Shock cells, causing cell damage. In addition, medium continuous perfusion culture was carried out using a feed pump 1 0 0 9 and a discharge pump 1 0 0 2 . The pressure-sensing valve 1 0 0 1 can also be turned on forever, and the timer (not shown) is used as the basis for the air pump 1 0 1 1 to operate. Or, the timer and the pressure-sensing valve member 1 0 0 1 are used together, that is, the pressure-sensing valve member 1 0 0 1 is pulse-opened and closed, so that the external chamber 1 0 0 4 pressure can be easily controlled, and The air pump 1 0 1 1 can also be turned off when the timer expires. The cell culture apparatus 100 of the present invention can be selectively operated by automation or by hand. Embodiment 2 The second A diagram and the second B diagram show a second embodiment of the present invention, a cell culture apparatus 200, which includes at least: an inner chamber 2 0 0 4; - an outer chamber 2 0 0 3 ; - filling structure 2 0 0 5, is disposed in the outer chamber 2 0 0 3 to fill the growth substrate 2008; - the upwardly inclined fin structure 2 0 0 9, is set in the aforementioned growth The substrate fills the structure of the wall edge to block the bubbles generated during the cultivation; the inner chamber 2004 and the outer chamber 2003 are in communication with each other. The culture device 200 further comprises: an inductive valve member 2 0 0 1 , an air pump 2 0 0 7 , a fresh medium container 2 0 1 0, a product collection container 2 0 1 1 , a feed pump 2 0 0 6 , discharge pump 2 0 0 2 . The cell culture device 200 can further selectively install an air filter 2 0 1 2 during the introduction of the air pump 2007 into the chamber to prevent contamination. The operation mode of the cell culture device 200 is as follows: as shown in the second A map
第19頁 1270576 五、發明說明（16) 示，當空氣幫浦2 0 0 7未做動前，大部分之培養基位於腔室 下層，因此生長基材2008暴露於空氣之中；如第二B圖所 示，當空氣幫浦2 0 0 7啟動後，培養基由内腔室2 0 0 4向外腔 室2 0 0 3壓縮，培養基上升於外腔室2 0 0 3而淹蓋住細胞生長 之生長基材填充結構2 0 0 5，當外腔室2 0 0 3壓力值上升至一 特定值之後，則關閉空氣幫浦2 0 0 7並將壓力感應式閥件 2 0 0 1打開，將外腔室2 0 0 3之壓力釋放，並使外腔室2 0 0 3之 培養基因重力流回内腔室2 0 0 4，此時生長基材2 0 0 8則又重 新暴露於空氣中，如第二A圖所示。如此反覆進行第二A圖 及第二B圖之程序，係可間歇性，但非直接的，經由一空 氣-培養基之薄層界面提供細胞氧氣，卻不會傷害及/或殺 死細胞。同時，為避免過量空氣吹入而導致培養基完全擠 入外腔室2003 ，並有多餘空氣由内腔室2004推入外腔室 2 0 0 3而造成氣泡直接衝擊生長基材2 0 0 8部分，因此利用一 設置於填充結構2 0 0 9壁緣之向上斜之鰭片結構2 0 0 9，將氣 泡導入外腔室2 0 0 3壁緣而阻絕氣泡之直接對細胞之衝擊。 此外，利用進料幫浦2 0 0 6與出料幫浦2 0 0 2進行培養基連續 灌注培養。另外，壓力感應式閥件2 0 0 1亦可永遠開啟，並 以計時器（圖未顯示）做為空氣幫浦2 0 0 7做動之依據，或 是，將計時器以及壓力感應式閥件2 0 0 1搭配使用，亦即壓 力感應閥件2 0 0 1採脈衝式啟閉，如此更可易於控制外腔室 2 0 0 3壓力，且當計時器時間到時亦可關閉空氣幫浦2 0 0 7。 本發明之細胞培養裝置2 0 0係可選擇性的以自動化或以手 動方式操作。Page 19 1270576 V. Description of the invention (16) shows that most of the medium is located in the lower layer of the chamber before the air pump 2 0 7 7 is not actuated, so the growth substrate 2008 is exposed to the air; As shown in the figure, when the air pump is activated, the medium is compressed by the inner chamber 2 0 0 4 to the outer chamber 2 0 0 3 , and the medium rises to the outer chamber 2 0 0 3 to flood the cell growth. The growth substrate filling structure 2 0 0 5, after the external chamber 2 0 0 3 pressure value rises to a specific value, the air pump 2 0 0 7 is closed and the pressure-sensitive valve member 2 0 0 1 is opened. The pressure of the outer chamber 2 0 0 3 is released, and the culture gene of the outer chamber 2 0 0 3 is gravity flowed back to the inner chamber 2 0 0 4 , at which time the growth substrate 2 0 0 8 is re-exposed to the air. Medium, as shown in Figure A. The procedure of Figure 2A and Figure 2B is repeated as such, intermittently, but not directly, to provide cellular oxygen through a thin layer interface of air-medium without harming and/or killing cells. At the same time, in order to avoid excessive air insufflation, the medium is completely squeezed into the outer chamber 2003, and excess air is pushed from the inner chamber 2004 into the outer chamber 2000, causing bubbles to directly impact the growth substrate 2 0 0 8 Therefore, by using an upwardly inclined fin structure 2 0 0 9 disposed on the wall edge of the filling structure 2 0 9 , the bubble is introduced into the outer chamber 2 0 0 3 wall edge to block the direct impact of the bubble on the cell. In addition, medium continuous perfusion culture was carried out using a feed pump 2 0 6 6 and a discharge pump 2 0 0 2 . In addition, the pressure-sensing valve member 2 0 0 1 can also be turned on forever, and the timer (not shown) is used as the basis for the air pump 2 0 0 7 or the timer and the pressure-sensing valve. The 2 0 0 1 is used together, that is, the pressure sensing valve member 2 0 0 1 is pulsed to open and close, so that the external chamber 2 0 0 3 pressure can be easily controlled, and the air can be turned off when the timer expires. Pu 2 0 0 7. The cell culture device 200 of the present invention can be selectively operated either automatically or manually.
第20頁 1270576 五、發明說明（17) 實施例三 第三A圖及第三B圖係顯示本發明之第三個實施例，一 種細胞培養裝置3 0 0，其至少包含：一内腔室3 0 0 3 ; —外 腔室3 0 0 4 ; —填充結構3 0 0 5，係設置於前述外腔室中，用 以填充生長基材3 0 0 8 ; —排氣通道3 0 0 9，係設置於前述内 腔室3 0 0 3頂部，以阻隔培養過程中產生之氣泡；前述内腔 室3 0 0 3與外腔室3 0 0 4之間係可相互連通。前述排氣通道 3 0 0 9下方進一步包含一浮板膈膜結構3 0 1 2，可將排氣通道 3 0 0 9阻斷，以防止培養基流出内腔3 0 0 3室而污染空氣濾網 3 0 1 3。此培養裝置3 0 0進一步包含：感應式閥件3 0 0 7、空 氣幫浦3 0 0 1 、新鮮培養基容器3 0 1 0、產物收集容器3 0 1 1 、 進料幫浦3 0 0 6、出料幫浦3 0 0 2。本細胞培養裝置3 0 0進一 步可於空氣幫浦3 0 0 1導入腔室過程中可以選擇性的裝設一 空氣濾網3 0 1 3以防止污染。 此細胞培養裝置3 0 0之操作方式如下：如第三A圖所 示，當空氣幫浦3 0 0 1未做動前，大部分之培養基位於腔室 下層，此時，生長基材3 0 0 8浸泡於培養基之中；如第三B 圖所示，當空氣幫浦3 0 0 1啟動後，培養基由外腔室3 0 0 4向 内腔室3 0 0 3壓縮而氣體則由内腔室3 0 0 3頂端之排氣通道 3 0 0 9排出，而培養基持續上升至當液面推動浮板膈膜結構 3012而將排氣通道3009阻斷時，外腔室3004壓力值上升至 一特定值之後則壓力感應閥件3 0 0 7便反饋控制空氣幫浦 3 0 0 1 ，將其關閉，並使内腔室3 0 0 3之培養基因重力流回外 腔室3 0 0 4，此時生長基材3 0 0 8則又重新暴露於空氣中，如Page 20 1270576 V. DESCRIPTION OF THE INVENTION (17) Embodiment 3 The third A diagram and the third B diagram show a third embodiment of the present invention, a cell culture apparatus 300, which includes at least: an inner chamber 3 0 0 3 ; - outer chamber 3 0 0 4 ; - filling structure 3 0 0 5, is disposed in the outer chamber to fill the growth substrate 3 0 0 8; - exhaust passage 3 0 0 9 The system is disposed at the top of the inner chamber 3 0 0 3 to block air bubbles generated during the cultivation process; the inner chamber 3 0 0 3 and the outer chamber 3 0 0 4 are in communication with each other. The exhaust passage 3 0 0 9 further includes a floating plate 膈 membrane structure 3 0 1 2, which can block the exhaust passage 3 0 0 9 to prevent the medium from flowing out of the inner chamber 3 0 0 3 chamber and pollute the air filter. 3 0 1 3. The culture device 300 further comprises: an inductive valve member 3 0 0 7 , an air pump 3 0 0 1 , a fresh medium container 3 0 1 0, a product collection container 3 0 1 1 , a feed pump 3 0 0 6 , discharge pump 3 0 0 2 . The cell culture device 300 can further selectively install an air filter 3 0 1 3 during the introduction of the air pump 300 into the chamber to prevent contamination. The operation mode of the cell culture device 300 is as follows: as shown in the third A figure, most of the culture medium is located in the lower layer of the chamber before the air pump 3 0 0 1 is not actuated. At this time, the growth substrate 3 0 0 8 immersed in the medium; as shown in the third B diagram, when the air pump 3000 starts, the medium is compressed from the outer chamber 3 0 0 4 to the inner chamber 3 0 0 3 and the gas is inside. The exhaust passage 3 0 0 9 at the top of the chamber 3 0 0 3 is discharged, and the medium continues to rise until the liquid level pushes the floating diaphragm structure 3012 to block the exhaust passage 3009, and the pressure of the outer chamber 3004 rises to After a certain value, the pressure sensing valve member 3 0 0 7 feeds back the control air pump 3 0 0 1 , closes it, and gravity flows the culture cell of the inner chamber 3 0 0 3 back to the outer chamber 3 0 0 4 At this time, the growth substrate 3 0 0 8 is re-exposed to the air, such as
第21頁 1270576 五、發明說明（18) 第三A圖所示。如此反覆進行第三A圖及第三B圖之程序， 係可間歇性，但非直接的，經由一空氣-培養基之薄層界 面提供細胞氧氣，卻不會傷害及/或殺死細胞。 此外，利用進料幫浦3 0 0 6與出料幫浦3 0 0 2進行培養基 連續灌注培養。本發明之細胞培養裝置3 0 0係可選擇性的 以自動化或以手動方式操作。 實施例四 第四A圖及第四B圖係顯示本發明之第四種實施例，一 種細胞培養裝置4 0 0，其至少包含：一内腔室4 0 0 4 ; —外 腔室4 0 0 3 ; —填充結構4 0 0 5，係設置於前述外腔室4 0 0 3 中，用以填充生長基材4 0 0 8 ; —壓力感應式閥件4 0 0 7，係 設置於前述内腔室4 0 0 4之頂部，以阻隔培養過程中產生之 氣泡；前述内腔室4 0 0 4與外腔室4 0 0 3之間係可相互連通。 此培養裝置4 0 0進一步包含：空氣幫浦4 0 0 1、新鮮培養基 容器4 0 1 0 、產物收集容器4 0 1 1 、進料幫浦4 0 0 6、出料幫浦 4002。本細胞培養裝置400進一步可於空氣幫浦4001導入 腔室過程中可以選擇性的裝設一空氣濾網4 0 0 9以防止污 染。 此細胞培養裝置4 0 0之操作方式如下：如第四A圖所 示，當空氣幫浦4 0 0 1未做動前，大部分之培養基位於腔室 下層，生長基材4 0 0 8浸泡於培養基之中；如第四B圖所 示，當空氣幫浦4 0 0 1啟動後，培養基由外腔室4 0 0 3向内腔 室4 0 04壓縮而内腔室4004氣體被壓縮，當内腔室4003壓力 值上升至一特定值之後，則壓力感應閥件4 0 0 7便反饋控制Page 21 1270576 V. Description of invention (18) Figure 3A shows. The procedure of the third and third panels is repeated in this manner to provide intermittent, but indirect, cellular oxygen through a thin layer of air-medium without harming and/or killing the cells. In addition, medium continuous perfusion culture was carried out using a feed pump 3 0 6 6 and a discharge pump 3000. The cell culture device 300 of the present invention can be selectively operated either automatically or manually. Embodiment 4 The fourth embodiment A and the fourth B show a fourth embodiment of the present invention, a cell culture device 400, which comprises at least: an inner chamber 4 0 0 4; - an outer chamber 40 0 3 ; - filling structure 4 0 0 5, is disposed in the outer chamber 4 0 0 3 to fill the growth substrate 4 0 0 8; - pressure-sensitive valve member 4 0 0 7, is set in the foregoing The top of the inner chamber 4 0 0 4 is used to block the bubbles generated during the cultivation process; the inner chamber 4 0 0 4 and the outer chamber 4 0 0 3 are in communication with each other. The culture device 400 further comprises: an air pump 4 0 0 1 , a fresh medium container 4 0 1 0 , a product collection container 4 0 1 1 , a feed pump 4 0 0 6 , a discharge pump 4002. The cell culture device 400 can further selectively install an air filter 4 0 0 9 during the introduction of the air pump 4001 into the chamber to prevent contamination. The operation mode of the cell culture device 400 is as follows: as shown in FIG. 4A, most of the culture medium is located in the lower layer of the chamber before the air pump 4 0 0 1 is not actuated, and the growth substrate is soaked in the 4 0 0 8 In the medium; as shown in FIG. 4B, when the air pump 4 0 0 1 is started, the medium is compressed from the outer chamber 400 to the inner chamber 410 and the inner chamber 4004 is compressed. When the pressure value of the inner chamber 4003 rises to a certain value, the pressure sensing valve member 4 0 0 7 is feedback controlled.
第22頁 1270576 五、發明說明（19) 空氣幫浦4 0 0 1 ，將其關閉，並使内腔室4 0 0 4之培養基因重 力流回外腔室4 0 0 3，此時生長基材4 0 0 8則又重新暴露於空 氣中，如第四A圖所示。如此反覆進行第四A圖及第四B圖 之程序，係可間歇性，但非直接的，經由一空氣-培養基 之薄層界面提供細胞氧氣，卻不會傷害及/或殺死細胞。 此外，利用進料幫浦4 0 0 6與出料幫浦4 0 0 2進行培養基連續 灌注培養。另外，壓力感應式閥件4 0 0 7亦可永遠開啟，並 以計時器（圖未顯示）做為空氣幫浦4 0 0 1做動之依據，或 是，將計時器以及壓力感應式閥件4 0 0 7搭配使用，亦即壓 力感應閥件4 0 0 7採脈衝式啟閉，如此更可易於控制外腔室 壓力，且當計時器時間到時亦可關閉空氣幫浦4 0 0 1 。 具體而言，本發明係關於一種可靠的、簡易的、不昂 貴的、可重複使用或可拋棄式的、無菌的與有效率的培養 細胞及/或組織與收集其產物之方法與裝置。更確切的 說，本發明係關於一新穎的方法與裝置可用來培養任何細 胞，不論是真核、原核、哺乳動物或動物細胞，並且可以 充分供應細胞生長所需之養分與氧氣卻不會傷害到細胞， 同時，藉由出口壓力感測與調控設備，可使培養腔室内的 壓力可以隨細胞之特性加以調整，以促使細胞生長或/與 分化。此外，本發明之方法與裝置可以預防或大量減低任 何形式的污染、避免造成剪應力與保護細胞不會直接接觸 空氣、氣泡或氣體。再者，本發明之裝置可為自動或手動 操作，可節省勞力及/或管理的人力，另外，本發明係提 供一較簡易、方便之方法與裝置，可用來生產及收集細胞Page 22 1270576 V. INSTRUCTIONS (19) The air pump 4 0 0 1 is closed, and the culture gene of the inner chamber 400 is gravity flowed back to the outer chamber 4 0 0 3 at this time. The material 4 0 0 8 is again re-exposed to the air, as shown in Figure 4A. The procedure of Figures 4A and 4B is repeated as such, intermittently, but not directly, to provide cellular oxygen through a thin layer interface of air-medium without harming and/or killing cells. In addition, the medium was continuously perfused with the feed pump 4 0 0 6 and the discharge pump 4 0 0 2 . In addition, the pressure-sensing valve member 4 0 0 7 can also be turned on forever, and the timer (not shown) is used as the basis for the air pump 4 0 0 1 or the timer and the pressure-sensing valve. The 4 0 0 7 is used together, that is, the pressure sensing valve member 4 0 0 7 is pulsed to open and close, so that the external chamber pressure can be easily controlled, and the air pump can be turned off when the timer expires. 1 . In particular, the present invention relates to a reliable, simple, inexpensive, reusable or disposable, sterile and efficient method of culturing cells and/or tissues and collecting products thereof. More specifically, the present invention relates to a novel method and apparatus for culturing any cell, whether eukaryotic, prokaryotic, mammalian or animal, and capable of adequately supplying the nutrients and oxygen required for cell growth without harming At the same time, by the outlet pressure sensing and regulating device, the pressure in the culture chamber can be adjusted according to the characteristics of the cells to promote cell growth or/and differentiation. Moreover, the method and apparatus of the present invention can prevent or substantially reduce any form of contamination, avoid shear stress and protect cells from direct contact with air, bubbles or gases. Furthermore, the apparatus of the present invention can be operated automatically or manually, saving labor and/or management manpower. In addition, the present invention provides a simple and convenient method and apparatus for producing and collecting cells.
第23頁 1270576 五、發明說明（20) 或組織的產物，例如：蛋白質、抗體等。 本發明係可藉由一較佳實施例來說明，需了解的是本 發明不應受限於所提供之實施態樣，而應包含各種本發明 之實施態樣（如同熟習本技術領域者所習知），因此，本 發明之申請專利範圍應作最廣義之解釋以包括所有修改與 相似的組合 本發明係利用一非限制性之實施例來進一步說明，本 實施例係用以描述本發明之内容，而不宜用來縮限本發明 及許多未脫離本發明精神與範疇之明顯的、可能的差異。Page 23 1270576 V. INSTRUCTIONS (20) or products of tissues, such as proteins, antibodies, etc. The present invention may be illustrated by a preferred embodiment, and it should be understood that the present invention should not be limited to the embodiments provided, but should include various embodiments of the present invention (as is apparent to those skilled in the art) The present invention is intended to be construed as broadly construed, The present invention is not intended to be used to limit the present invention and many obvious and possible differences without departing from the spirit and scope of the invention.
第24頁 P^70576 圖式簡單說明 第一 A圖係顯示本發明之細胞培養裝置之第一個實施 例，其中空氣幫浦未做動前，生長基材係暴露於空氣中之 狀態。 第一 B圖係顯示本發明之細胞培養裝置之第一個實施 例，其中空氣幫浦做動後，生長基材浸泡於培養基中之狀 態。 第二A圖係顯示本發明之細胞培養裝置之第二個實施 例，其中空氣幫浦未做動前’生長基材係暴露於空氣中之 狀態。Page 24 P^70576 Brief Description of the Drawing The first A diagram shows the first embodiment of the cell culture apparatus of the present invention in which the growth substrate is exposed to the air before the air pump is actuated. The first B diagram shows the first embodiment of the cell culture apparatus of the present invention, in which the growth substrate is immersed in the medium after the air pump is actuated. Fig. 2A shows a second embodiment of the cell culture apparatus of the present invention in which the air pump is not exposed to the state in which the growth substrate is exposed to the air.
第二B圖係顯示本發明之細胞培養裝置之第二個實施 例，其中空氣幫浦做動後，生長基材浸泡於培養基中之狀 態。 第三A圖係顯示本發明之細胞培養裝置之第三個實施 例，其中空氣幫浦未做動前，生長基材浸泡於培養基中之 狀態。 第三B圖係顯示本發明之細胞培養裝置之第三個實施 例，其中空氣幫浦做動後，生長基材係暴露於空氣中之狀 態。Fig. 2B shows a second embodiment of the cell culture apparatus of the present invention, in which the growth substrate is immersed in the medium after the air pump is actuated. Fig. 3A shows a third embodiment of the cell culture apparatus of the present invention, in which the growth substrate is immersed in the medium before the air pump is actuated. Fig. 3B shows a third embodiment of the cell culture apparatus of the present invention, in which the growth substrate is exposed to the air after the air pump is actuated.
第四A圖係顯示本發明之細胞培養裝置之第四個實施 例，其中空氣幫浦未做動前，生長基材浸泡於培養基中之 狀態。 第四B圖係顯示本發明之細胞培養裝置之第四個實施 例，其中空氣幫浦做動後，生長基材係暴露於空氣中之狀 態0Fig. 4A shows a fourth embodiment of the cell culture apparatus of the present invention, in which the growth substrate is immersed in the medium before the air pump is actuated. Fig. 4B is a view showing a fourth embodiment of the cell culture apparatus of the present invention, in which the growth substrate is exposed to the air state after the air pump is actuated.
第25頁 1270576 圖式簡單說明 【主要原件符號對照說明】 1 〇〇-- -細胞培養裝置 100 1- ——壓 力 感 應 閥件 1 0 0 2 - —出 料 幫 浦 1 0 0 3 - —產 物 收 集 容器 1 0 0 4- 一一外 腔 室 1 0 0 5 - 一一内 腔 室 1 0 0 6 - --階 梯 式 斜 板結 構 1 0 0 7 - --填 充 結 構 1 0 0 8 - --新 鮮 培 養 基容 器 1 0 0 9 - ——進 料 幫 浦 1010 - --生 長 基 材 ion - --空 氣 幫 浦 10 12- __空 氣 濾 網 2 0 0 -- -細胞培養裝置 2 0 0 1 - --壓 力 感 應 式閥 件 2 0 0 2 - —出 料 幫 浦 2 0 0 3 - —外 腔 室 2 0 0 4 - ——内 腔 室 2 0 0 5 - --填 充 結 構 2 0 0 6 - ——進 料 幫 浦 2 0 0 7 - 一一空 氣 幫 浦 2 0 0 8 - --生 長 基 材 2 0 0 9 - —彥耆 片 式 結 構Page 25 1270576 Brief description of the diagram [Main original symbol comparison description] 1 〇〇-- - Cell culture device 100 1- —— Pressure sensing valve member 1 0 0 2 - - Discharge pump 1 0 0 3 - Product Collection container 1 0 0 4- one outer chamber 1 0 0 5 - one inner chamber 1 0 0 6 - - stepped sloping plate structure 1 0 0 7 - --fill structure 1 0 0 8 - -- Fresh medium container 1 0 0 9 - - Feed pump 1010 - - Growth substrate ion - - Air pump 10 12 - __ Air filter 2 0 0 -- - Cell culture device 2 0 0 1 - -- Pressure-sensitive valve member 2 0 0 2 - - Discharge pump 2 0 0 3 - - External chamber 2 0 0 4 - - Inner chamber 2 0 0 5 - - Filled structure 2 0 0 6 - ——Feeding pump 2 0 0 7 - One air pump 2 0 0 8 - --Growing substrate 2 0 0 9 -—Yan Yan film structure
第26頁 1270576Page 26 1270576
第27頁 圖式簡單說明 20 10- --新鮮培養基容 20 11- --產物收集容器 2012- —空氣渡網 3 0 0 -- -細胞培養裝置 3 0 0 1 - --空氣幫浦 3 0 0 2 - --出料幫浦 3 0 0 3 - --内腔室 3 0 0 4 - --外腔室 3 0 0 5 - —填充結構 3 0 0 6 - --進料幫浦 3 0 0 7 - - -壓力感應式閥 3 0 0 8 - —生長基材 3 0 0 9 - --排氣通道 30 10- —新鮮培養基容 30 11- --產物收集容器 3012- —浮板腺膜 30 13- --空氣濾網 4 0 0 — -細胞培養裝置 4 0 0 1 - —空氣幫浦 4 0 0 2 - --出料幫浦 4 0 0 3 - --外腔室 4 0 0 4- --内腔室 4 0 0 5 - —填充結構 4 0 0 6 - --進料幫浦 1270576 圖式簡單說明 4 0 0 7 ---壓力感應式閥件 4 0 0 8 ---生長基材 4 0 0 9——空氣濾網 4 0 10---新鮮培養基容器 4 0 1 1 - --產物收集容器Page 27 Schematic description 20 10 -- Fresh medium capacity 20 11- -- Product collection container 2012 - Air network 3 0 0 -- - Cell culture device 3 0 0 1 - Air pump 3 0 0 2 - -- Discharge pump 3 0 0 3 - -- Internal chamber 3 0 0 4 - - External chamber 3 0 0 5 - Filling structure 3 0 0 6 - - Feeding pump 3 0 0 7 - - - Pressure-sensitive valve 3 0 0 8 - - Growth substrate 3 0 0 9 - - Exhaust channel 30 10- - Fresh medium volume 30 11- -- Product collection container 3012 - Floating plate gland 30 13- -- Air filter 4 0 0 — - Cell culture device 4 0 0 1 - Air pump 4 0 0 2 - -- Discharge pump 4 0 0 3 - -- External chamber 4 0 0 4 - - Inner chamber 4 0 0 5 - Filling structure 4 0 0 6 - - Feeding pump 1270576 Schematic description 4 0 0 7 --- Pressure sensing valve 4 0 0 8 --- Growth Substrate 4 0 0 9 - air filter 4 0 10---fresh medium container 4 0 1 1 - -- product collection container
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| Application Number | Priority Date | Filing Date | Title |
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| TW92112990ATWI270576B (en) | 2003-05-13 | 2003-05-13 | Cell-cultivating device and method |
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| TWI270576Btrue TWI270576B (en) | 2007-01-11 |
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| TW92112990ATWI270576B (en) | 2003-05-13 | 2003-05-13 | Cell-cultivating device and method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US10704017B2 (en) | 2017-02-03 | 2020-07-07 | Industrial Technology Research Institute | Cell culture carrier module and cell culture system |
| US11884912B2 (en) | 2018-12-27 | 2024-01-30 | Industrial Technology Research Institute | Cell culture device and method thereof |
| TWI852134B (en)* | 2022-10-19 | 2024-08-11 | 緯創資通股份有限公司 | Vertical transfer equipment |
| TWI863949B (en)* | 2018-12-28 | 2024-12-01 | 日商昕芙旎雅股份有限公司 | Cell culture systems and cell culture devices |
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| US20210371788A1 (en)* | 2020-05-26 | 2021-12-02 | Cesco Bioengineering Co., Ltd. | Apparatus and method for culturing cells |
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| US10704017B2 (en) | 2017-02-03 | 2020-07-07 | Industrial Technology Research Institute | Cell culture carrier module and cell culture system |
| US11884912B2 (en) | 2018-12-27 | 2024-01-30 | Industrial Technology Research Institute | Cell culture device and method thereof |
| TWI863949B (en)* | 2018-12-28 | 2024-12-01 | 日商昕芙旎雅股份有限公司 | Cell culture systems and cell culture devices |
| TWI852134B (en)* | 2022-10-19 | 2024-08-11 | 緯創資通股份有限公司 | Vertical transfer equipment |
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| TW200424311A (en) | 2004-11-16 |
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