Cell culture device capable of synchronously replacing culture mediumTechnical Field
The invention relates to the technical field of microorganism culture, in particular to a cell culture device capable of synchronously replacing culture mediums.
Background
As is well known, a device capable of simultaneously culturing a large number of cells is typically a well plate (or microplate), and specifically, the well plate includes a plate body and a large number of base wells opened on the plate body, a culture medium is injected into each of the base wells, and cells to be cultured are implanted into the base wells, whereby a large number of cells can be simultaneously cultured using the large number of base wells of the well plate.
The prior art has the defects that the base holes are independent from each other, when the culture medium is required to be replaced, the culture medium is sucked and injected into each base hole simultaneously by using equipment with a plurality of needle tubes which are arranged in line with the base holes, and the culture medium is sucked and injected into each base hole independently and sequentially by using one needle tube, so that the culture medium replacement of the orifice plate with the structure is finished by using special and expensive equipment, and the orifice plate is required to be placed in the environment of the equipment. In addition, in some cases, for example, if the volume of cells is small, when the medium is replaced by aspiration with a needle, the cells may be aspirated into the needle simultaneously, with undesirable removal consequences for the cells.
To overcome the above-mentioned drawbacks, a well plate for conveniently replacing (or transferring) a culture medium has been developed in the prior art, and in particular, the well plate is improved over the conventional well plate in that each column (row) of base wells arranged in a matrix is communicated by means of fluid channels, i.e. fluid channels penetrating two base wells are machined between each adjacent two base wells of each column (row). In this way, the effect of removing the medium can be obtained by sucking the medium from the tail of each column (row) or tilting the well plate so that the medium in each base well flows to the tail, and the medium can be replaced by injecting the medium into the base well of the head so that the medium is gradually injected into each medium.
However, the improved well plate of the prior art still has the disadvantage during use that during removal of the medium by means of the fluid channel and injection of the medium into the base wells, some of the cells in the base wells may migrate with the medium to other base wells and even out of the well plate with the medium, thereby causing undesired removal consequences for the cells.
Disclosure of Invention
Aiming at the technical problems in the prior art, the invention provides a cell culture device capable of synchronously replacing culture mediums.
In order to solve the technical problems, the invention adopts the following technical scheme:
a cell culture apparatus with synchronized medium exchange, comprising:
The box main body comprises a rectangular plate body and coamings arranged around the side edges of the plate body, wherein the coamings and the upper surface of the plate body enclose a main body pool capable of containing culture mediums;
the base holes are formed in the upper plate surface of the plate body, and the base holes comprise a plurality of base holes which are arranged in a matrix manner, so that the culture medium flows along the row arrangement direction of the base holes;
The filter cartridges are arranged above each base hole, downstream overflow gaps are formed in the walls of the filter cartridges, which are located on one downstream side at least, of the filter cartridges, and the filter cartridges are characterized in that:
a sinking cavity is formed in the upper plate surface of the plate body at the upstream side of each base hole, an upstream hole wall is defined between the sinking cavity and the base hole, and a first upstream overflow gap is formed in the upstream hole wall;
A water blocking dam is arranged between every two adjacent filter cartridges of each row of filter cartridges, the main body tanks are divided into tank units by the water blocking dams and the filter cartridges of the adjacent rows, and each row of water blocking dam enables culture medium in the tank units to actively flow to the sinking cavity and enter the base holes through the first upstream overflow gap.
Preferably, the through-flow cross section defined by the downstream flow gap through which the culture medium is allowed to pass is larger than the through-flow cross section defined by the first upstream flow gap through which the culture medium is allowed to pass.
Preferably, a second upstream flow gap is formed in the wall of the filter cartridge on the upstream side, and the through-flow section defined by the second upstream flow gap for allowing the culture medium to pass through is smaller than the through-flow section defined by the first upstream flow gap for allowing the culture medium to pass through.
Preferably, the sum of the through-flow cross sections of the first upstream flow gap and the second upstream flow gap is smaller than the through-flow cross section of the downstream flow gap.
Preferably, the width of the first upstream flow gap increases gradually from top to bottom, and the lower end of the first upstream flow gap is configured as a passivated arc.
Preferably, the height of the water retaining dam of each row is higher than the height of the filter cartridge of the row.
Preferably, the check dams arranged in a matrix and the filter cartridges arranged in a matrix are integrally formed by plastic materials through injection molding or laser etching processes.
Preferably, the cell culture device capable of synchronously replacing the culture medium further comprises a transferable component, the transferable component comprises a mesh body and a plurality of culture baskets, the mesh body is used for being attached to the upper surface of the plate body, the mesh body is provided with hollowed-out parts corresponding to the base holes one by one, the culture baskets are correspondingly arranged below the hollowed-out parts and are attached to the mesh body, the culture baskets extend into the base holes, and the cells are located in the culture baskets.
Preferably, the two opposite coamings are respectively provided with a liquid inlet port and a liquid outlet port.
Preferably, the upper portion of the cartridge body is detachably covered with a transparent cover.
Compared with the prior art, the cell culture device capable of synchronously replacing the culture medium has the beneficial effects that:
By arranging the interception components of the water retaining dams and the filter cartridges alternately in the main body pool of the box main body, arranging the sinking cavity at the upstream of each base hole, and arranging the downstream gap on the downstream cylinder wall of the filter cartridge and the upstream gap on the upstream hole wall of the base hole, the culture medium is forced to flow downwards preferentially and enter the base holes, and the replacement efficiency of the culture medium around the cells can be remarkably improved on the premise that the cells cannot generate undesired migration.
An overview of various implementations or examples of the technology described in this disclosure is not a comprehensive disclosure of the full scope or all of the features of the technology disclosed.
Drawings
In the drawings, which are not necessarily drawn to scale, like numerals may describe similar components in different views. The same reference numerals with letter suffixes or different letter suffixes may represent different instances of similar components. The accompanying drawings illustrate various embodiments by way of example in general and not by way of limitation, and together with the description and claims serve to explain the inventive embodiments. Wherever possible, the same reference numbers will be used throughout the drawings to refer to the same or like parts. Such embodiments are illustrative and not intended to be exhaustive or exclusive of the present apparatus or method.
FIG. 1 is a top view of a cell culture apparatus capable of synchronously changing culture medium according to the present invention.
Fig. 2 is an enlarged view of a portion B of fig. 1.
Fig. 3 is a cross-sectional view taken along A-A of fig. 1.
Fig. 4 is an enlarged view of a portion C of fig. 3.
Reference numerals:
10-box main body, 20-plate body, 21-base hole, 22-upstream hole wall, 23-sinking cavity, 31-back panel, 32-front panel, 33-side panel, 34-main body tank, 35-tank unit, 40-filter cartridge, 50-retaining dam, 61-downstream gap, 62-first upstream gap, 63-second upstream gap, 71-liquid inlet interface, 72-liquid outlet interface, 80-transparent cover body, 90-transferable component, 91-net sheet body and 92-culture basket.
Detailed Description
Unless defined otherwise, technical or scientific terms used herein should be given the ordinary meaning as understood by one of ordinary skill in the art to which this invention belongs. The terms "first," "second," and the like, as used herein, do not denote any order, quantity, or importance, but rather are used to distinguish one element from another. The word "comprising" or "comprises", and the like, means that elements or items preceding the word are included in the element or item listed after the word and equivalents thereof, but does not exclude other elements or items. The terms "connected" or "connected," and the like, are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect. "upper", "lower", "left", "right", etc. are used merely to indicate relative positional relationships, which may also be changed when the absolute position of the object to be described is changed.
In order to keep the following description of the embodiments of the present invention clear and concise, the detailed description of known functions and known components thereof have been omitted.
As shown in fig. 1 to 4, the present invention discloses a cell culture apparatus capable of simultaneously culturing a large number of cells by disposing a plurality of base holes 21, comprising a cartridge body 10, a transferable member 90, an intercepting member, and a transparent cover 80.
The box main body 10 includes a rectangular plate body 20, and a dash panel 32, a back panel 31 and two side panels 33 arranged around four sides of the plate body 20, the dash panel 32, the back panel 31, the two side panels 33 and the upper surface of the plate body 20 enclose together a main body pool 34 in which a culture medium can be accommodated, a liquid inlet port 71 communicating with the main body pool 34 is arranged on the back panel 31 (or called upstream panel), and a liquid outlet port 72 communicating with the main body pool 34 is arranged on the dash panel 32 (or called downstream panel). When the medium needs to be replaced, a new medium is made to enter the main body tank 34 via the liquid inlet port 71, and then flows out of the liquid outlet port 72 after passing through the main body tank 34.
The plate body 20 may be made of a biocompatible material, for example, a biocompatible polymer through injection molding, or may be made of a biocompatible alloy material, for example, a titanium alloy, and preferably, the plate body 20 is made of a biocompatible polymer through injection molding.
The upper surface of the plate body 20 is provided with a plurality of base holes 21, the plurality of base holes 21 are arranged in a matrix in a plurality of rows and columns, the base holes 21 are arranged in a row manner in the direction of the front coaming 32 and the rear coaming 31, and new culture medium sequentially passes through the base holes 21 of each row. The upper plate surface of the plate body 20 on the upstream side of each base hole 21 of each row is provided with a sinking cavity 23, and the sinking cavity 23 does not penetrate through the base holes 21, but together with the base holes 21, defines an upstream hole wall 22. The base hole 21 and the sinking cavity 23 can be integrally injection molded with the plate body 20, or can be molded by engraving.
The upstream wall 22 of the base aperture 21 is provided with a plurality of first upstream flow slits 62, which allow the medium entering the submerged chamber 23 to enter the base aperture 21 through the first upstream flow slits 62. Preferably, the first upstream flow gap 62 may be shaped by engraving. Preferably, the width of the first upstream flow gap 62 increases gradually from top to bottom, which makes it easier for the culture medium to enter the bottom of the base well 21 from the lower portion of the upstream well wall 22. More preferably, the lower end of the first upstream flow gap 62 is machined with an arcuate passivation to avoid clogging of this area with impurities.
The interception member includes a plurality of filter cartridges 40 and a water blocking dam 50, each of which has the filter cartridges 40 above each of the base holes 21, such that the filter cartridges 40 are also arranged in a matrix of a plurality of rows and columns, the water blocking dam 50 is disposed between each adjacent two of the filter cartridges 40 of each row, such that the filter cartridges 40 of each row are alternately arranged with the water blocking dam 50, and the pool unit 35 is defined between each adjacent two rows of the filter cartridges 40 and the water blocking dam 50, such that the main body pool 34 is divided into a plurality of pool units 35 arranged in the flow direction of the medium. When it is desired to replace the medium, the filter cartridge 40 and the retaining dam 50 located in the same row have an intercepting effect on the medium upstream thereof to force the medium into the sinking chamber 23 corresponding to each of the base holes 21 and into the base holes 21 via the first upstream flow gap 62 of the upstream hole wall 22 for replacement of the medium in the base holes 21.
The wall of the downstream side of each filter cartridge 40 is provided with a plurality of downstream flow-through slits 61, and obviously, the height of the downstream flow-through slits 61 is higher than that of the first upstream flow-through slits 62, so that the culture medium entering the base holes 21 through the first upstream flow-through slits 62 flows out of the downstream flow-through slits 61 after upward movement, and the cells in the base holes 21 are intercepted by the wall of the filter cartridge 40 without undesired migration, and thus, the replacement speed of the culture medium in the base holes 21 is obviously increased without undesired migration of the cells. Preferably, the matrix of filter cartridges 40 and the matrix of water dams 50 are integrally injection molded from a plastic material so that the interception member can be removably mounted in the body sump 34, which facilitates cleaning of the body sump 34 of the cartridge body 10.
The transferable member 90 includes a mesh body 91 and a plurality of culture baskets 92 arranged in a matrix attached to the bottom of the mesh body 91, the mesh body 91 is attached to the upper surface of the plate body 20, the mesh body 91 has hollowed-out portions corresponding to the base holes 21 one by one, and the culture baskets 92 are attached to the bottoms of the hollowed-out portions, so that the culture baskets 92 are located in the base holes 21, and cells are placed in the culture baskets 92 for culture. When it is desired to transfer the cells in each of the wells 21 in its entirety, the interceptor member is first removed and then the entire transfer of the cells can be accomplished by transferring the transferable member 90.
A transparent cover 80 is provided to cover the cartridge body 10 over each base hole 21, and the transparent cover 80 may be made of inorganic glass or organic glass. The transparent cover 80 can block the interference of the external air and the impurities thereof and can facilitate the observation of the growth of the cells in each of the base holes 21.
In some preferred embodiments, a plurality of second upstream flow-through slits 63 are provided in the wall of the cartridge on the upstream side of each filter cartridge 40, which allows a portion of the upstream medium to pass through the second upstream flow-through slits 63 to the area above the base aperture 21 for replacement of the medium above the base aperture 21, except for a portion of the upstream medium which passes through the sinking chamber 23 and the first upstream flow-through slits 62 to the middle lower portion of the base aperture 21.
In some preferred embodiments, the through-flow cross-section through which the culture medium is allowed to pass, which is defined by all of the first upstream flow-through slits 62, is made larger than the through-flow cross-section through which the culture medium is allowed to pass, which is defined by all of the second upstream flow-through slits 63, which allows more culture medium to pass through the first upstream flow-through slits 62 into the base aperture 21, thereby allowing the culture medium around the cells to be replaced more efficiently.
In some preferred embodiments, the sum of the through-flow cross-sections of all the first upstream flow-through slits 62 and all the second upstream flow-through slits 63 is made smaller than the through-flow cross-section of all the downstream flow-through slits 61, so that the level of the medium in the filter cartridge 40 is always smaller than the level of the medium upstream of the flow-through cartridge when the medium is replaced, thereby further improving the efficiency of replacement of the base well 21 and the medium above it.
In some preferred embodiments, the height of the retaining dam 50 is greater than the height of the filter cartridge 40, such that the medium in each pond unit 35 flows only through the base aperture 21 to the next pond unit 35.
The culture device disclosed by the invention has at least the following advantages:
By providing the intercepting members in which the check dams 50 are alternately arranged with the filter cartridges 40 in the body sump 34 of the cartridge body 10, and by providing the sinking chamber 23 upstream of each of the base holes 21 and providing the downstream slit on the downstream cylinder wall of the filter cartridge 40 and the upstream slit on the upstream cylinder wall 22 of the base hole 21, the culture medium is forced to flow downward preferentially into the base holes 21, whereby the exchange efficiency of the culture medium around the cells can be remarkably improved without causing undesired migration of the cells.
Furthermore, although exemplary embodiments have been described in the present disclosure, the scope thereof includes any and all embodiments having equivalent elements, modifications, omissions, combinations (e.g., of the various embodiments across), adaptations or alterations as would be appreciated by those in the art. The elements in the claims are to be construed broadly based on the language employed in the claims and are not limited to examples described in the present specification or during the practice of the application, which examples are to be construed as non-exclusive. It is intended, therefore, that the specification and examples be considered as exemplary only, with a true scope and spirit being indicated by the following claims and their full scope of equivalents.
The above description is intended to be illustrative and not restrictive. For example, the above-described examples (or one or more aspects thereof) may be used in combination with each other. For example, other embodiments may be used by those of ordinary skill in the art upon reading the above description. In addition, in the above detailed description, various features may be grouped together to streamline the invention. This is not to be interpreted as an intention that the disclosed features not being claimed are essential to any claim. Rather, inventive subject matter may lie in less than all features of a particular disclosed embodiment. Thus, the following claims are hereby incorporated into the detailed description as examples or embodiments, with each claim standing on its own as a separate embodiment, and it is contemplated that these embodiments may be combined with one another in various combinations or permutations. The scope of the invention should be determined with reference to the appended claims, along with the full scope of equivalents to which such claims are entitled.
The above embodiments are only exemplary embodiments of the present invention and are not intended to limit the present invention, the scope of which is defined by the claims. Various modifications and equivalent arrangements of this invention will occur to those skilled in the art, and are intended to be within the spirit and scope of the invention.