Cell culture device

Abstract

The invention relates to the technical field of microbial culture, and discloses a cell culture device, which comprises: a plate body; the culture units are arranged on the plate body at intervals in the width direction of the plate body, and each culture unit comprises a plurality of base holes which are arranged at intervals in the length direction of the plate body and are used for culturing cells and fluid channels which are arranged between every two adjacent base holes; according to the flowing direction of the culture medium along the fluid channels, the two sections of fluid channels corresponding to the two upward sides of each base hole are called the upstream fluid channel and the downstream fluid channel of the base hole; a downstream hole wall is arranged at least between the fluid channel at the downstream of the base hole and the base hole, and a plurality of downstream gaps allowing the culture medium to pass through and limiting the cells to pass through are formed in the downstream hole wall in a cutting manner.

Description

Technical Field

[0001] The invention relates to the technical field of microbial culture, and in particular to a cell culture device. Background Art

[0002] As is known, a typical device capable of culturing a large number of cells simultaneously is a well plate (or microwell plate). Specifically, the well plate includes a plate body and a large number of base holes opened on the plate body. Culture medium is injected into each base hole, and cells to be cultured are implanted into the base holes. Thus, a large number of cells can be cultured simultaneously using the numerous base holes of the well plate.

[0003] The above-mentioned orifice plate in the prior art has the following defects in culturing cells: the base holes are independent of each other. When the culture medium needs to be replaced, a device with a plurality of needles arranged in accordance with the base holes is used to simultaneously aspirate and inject the culture medium into each base hole. Otherwise, a needle is required to aspirate and inject the culture medium into each base hole in sequence. Therefore, the orifice plate of the above structure needs to use special and expensive equipment to complete the replacement of the culture medium, and the orifice plate needs to be placed in the environment where the equipment is located. In addition, in some cases, for example, if the cell volume is small, when the culture medium is replaced by aspiration using a needle, the cell may be sucked into the needle at the same time, resulting in the undesirable removal of the cell.

[0004] In order to overcome the above-mentioned defects, a well plate that is convenient for replacing (or transferring) culture medium has appeared in the prior art. Specifically, the improvement of the well plate relative to the traditional well plate is that each column (row) of the matrix-arranged basic holes is connected by a fluid channel, that is, a fluid channel that passes through the two basic holes is processed between each two adjacent basic holes in each column (row). In this way, the culture medium can be removed by aspirating the culture medium from the tail of each column (row) or by tilting the well plate so that the culture medium in each basic hole flows to the tail, and the culture medium can be gradually injected into each culture medium by injecting the culture medium into the basic hole at the head, thereby realizing the replacement of the culture medium.

[0005] However, the improved orifice plate in the prior art still has the following defects during use: in the process of removing the culture medium by the fluid channel and injecting the culture medium into the base well, some cells in the base well may be transferred to other base wells with the culture medium, or even flow out of the plate well with the culture medium, thereby causing undesirable removal of cells. Although technicians have tried to prevent cell transfer by implanting filter nets in the base wells, these methods are ultimately difficult to implement because the size of the base wells is basically in the millimeter level or even in the micrometer level, and the filter nets adapted to this size are difficult to prepare or install. Summary of the invention

[0006] In view of the above technical problems existing in the prior art, the present invention provides a cell culture device.

[0007] In order to solve the above technical problems, the technical solution adopted by the present invention is: A cell culture device, comprising: plate body; A culture unit, comprising a plurality of culture units, which are arranged on the plate body at intervals along the width direction of the plate body, each culture unit comprising a plurality of base holes for culturing cells arranged at intervals along the length direction of the plate body and a fluid channel between each two adjacent base holes, and according to the flow direction of the culture medium along the fluid channel, the two sections of the fluid channel corresponding to the two sides of each base hole in the radial direction are called the fluid channel upstream of the base hole and the fluid channel downstream of the base hole; wherein: A downstream hole wall is configured between at least the fluid channel downstream of the basic hole and the basic hole, and a plurality of downstream gaps that allow the medium to pass through but restrict the cells to pass through are processed on the downstream hole wall by cutting.

[0008] Preferably, an upstream pore wall is arranged between the fluid channel upstream of the base pore and the base pore, and a plurality of upstream slits that allow the medium to pass through but restrict the cells to pass through are processed on the upstream pore wall by cutting.

[0009] Preferably, A downstream flow-increasing cavity extending in the circumferential direction is arranged between the downstream fluid channel of the base hole and the downstream hole wall, the downstream fluid channel of the base hole passes through the downstream flow-increasing cavity, the width of the downstream flow-increasing cavity is greater than the width of the port of the downstream fluid channel of the base hole so that the width of the downstream hole wall is greater than the width of the port of the downstream fluid channel of the base hole; An upstream flow-increasing cavity extending in the circumferential direction is arranged between the fluid channel upstream of the base hole and the upstream hole wall, the fluid channel upstream of the base hole passes through the upstream flow-increasing cavity, the width of the upstream flow-increasing cavity is greater than the width of the port of the fluid channel upstream of the base hole so that the width of the upstream hole wall is greater than the width of the port of the fluid channel upstream of the base hole.

[0010] Preferably, all the base holes of each culture unit are located in the same plane; the bottom of the upstream flow-increasing cavity of each base hole is lower than the bottom of the downstream flow-increasing cavity, and the bottom of the fluid channel between two adjacent base holes is an inclined surface, and the two ends of the inclined surface are respectively connected to the bottom of the downstream flow-increasing cavity and the bottom of the upstream flow-increasing cavity; wherein: the bottom of the upstream gap is not higher than the bottom of the corresponding upstream flow-increasing cavity, and the bottom of the downstream gap is not lower than the bottom of the corresponding downstream flow-increasing cavity.

[0011] Preferably, the base hole comprises a circular hole section located at the upper portion and a tapered hole section located at the lower portion.

[0012] Preferably, the bottom of the downstream gap is located in the circular hole section, and the bottom of the upstream gap is located in the tapered hole section.

[0013] Preferably, the culture unit further comprises an upstream liquid storage tank located upstream of the most upstream base hole and a downstream liquid storage tank located downstream of the most downstream base hole.

[0014] Preferably, the two upstream liquid storage tanks and the two downstream liquid storage tanks of two adjacent culture units are separated by a partition; wherein: A notch extending downward from the top is formed on each of the partitions, and a blocking component is detachably mounted on the notch.

[0015] Preferably, two end surfaces in the length direction of the plate body are provided with liquid inlet interfaces corresponding one-to-one to the plurality of upstream liquid storage tanks and liquid outlet interfaces corresponding one-to-one to the plurality of downstream liquid storage tanks.

[0016] Preferably, the upstream hole wall and the upstream slit on the upstream hole wall, the downstream hole wall and the downstream slit on the downstream hole wall are all obtained by laser cutting.

[0017] Compared with the prior art, the cell culture device provided by the present invention has the following beneficial effects: The present invention configures a pore wall with a gap between the fluid channel and the base pore, so that during the replacement of the culture medium or the recycling of the culture medium, the pore wall allows the culture channel to restrict the passage of cells, thereby avoiding the undesirable transfer of cells.

[0018] This summary of various implementations or examples of the technology described in this disclosure is not a comprehensive disclosure of the entire scope or all features of the disclosed technology. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In the drawings, which are not necessarily drawn to scale, the same reference numerals may describe similar parts in different views. The same reference numerals with letter suffixes or different letter suffixes may represent different instances of similar parts. The accompanying drawings generally illustrate various embodiments by way of example and not limitation, and together with the specification and claims, are used to illustrate the embodiments of the invention. When appropriate, the same reference numerals are used throughout the drawings to refer to the same or similar parts. Such embodiments are illustrative and are not intended to be exhaustive or exclusive embodiments of the present apparatus or method.

[0020] Figure 1 This is a schematic diagram of the three-dimensional structure of the cell culture device provided by the present invention.

[0021] Figure 2 for Figure 1 An enlarged view of detail A.

[0022] Figure 3 This is a top view of the cell culture device provided by the present invention.

[0023] Figure 4 for Figure 3 An enlarged view of detail B.

[0024] Figure 5 This is a three-dimensional cross-sectional view of the cell culture device provided by the present invention.

[0025] Figure 6 for Figure 5 An enlarged view of a portion C.

[0026] Figure 7 This is a schematic structural diagram of a blocking component in the cell culture device provided by the present invention.

[0027] Reference numerals: 10-plate body; 20-culture unit; 21-base hole; 22-fluid channel; 23-upstream liquid storage tank; 231-liquid inlet interface; 232-upstream partition; 233-upstream gap; 24-downstream liquid storage tank; 241-liquid outlet interface; 242-downstream partition; 243-downstream gap; 25-sealing component. DETAILED DESCRIPTION

[0028] Unless otherwise defined, the technical terms or scientific terms used in the present invention should be understood by people with ordinary skills in the field to which the present invention belongs. The words "first", "second" and similar words used in the present invention do not indicate any order, quantity or importance, but are only used to distinguish different components. "Include" or "comprise" and similar words mean that the elements or objects appearing before the word include the elements or objects listed after the word and their equivalents, without excluding other elements or objects. "Connect" or "connected" and similar words are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect. "Up", "down", "left", "right" and the like are only used to indicate relative positional relationships. When the absolute position of the described object changes, the relative positional relationship may also change accordingly.

[0029] In order to keep the following description of the embodiments of the present invention clear and concise, detailed descriptions of well-known functions and well-known components are omitted.

[0030] like Figures 1 to 7 As shown, the present invention discloses a cell culture device, which is used to culture a large number of cells at the same time. The device includes a plate body 10 and a plurality of culture units 20 arranged on the upper surface of the plate body 10 .

[0031] The plate body 10 can be made of a biocompatible material, for example, a biocompatible high molecular polymer by injection molding, or a biocompatible alloy material, for example, a titanium alloy, or a composite of a titanium alloy and a high molecular polymer. Preferably, the plate body 10 is made of a titanium alloy or a composite of a titanium alloy on the upper part and a high molecular polymer on the lower part.

[0032] like Figures 1 to 4 As shown, the culture unit 20 includes a plurality of culture units 20, which are arranged at intervals along the width direction of the plate body 10 and are configured on the upper part of the plate body 10. Each culture unit 20 includes a plurality of base holes 21 arranged at intervals along the length direction of the plate body 10 and a fluid channel 22 between every two base holes 21, and the fluid channel 22 allows the base holes 21 to be interconnected to allow fluid to flow. Each base hole 21 is used to culture cells, so each base hole 21 can be implanted with cells, and the cells are cultured by injecting culture medium into the base hole 21.

[0033] Since each adjacent two base holes 21 are connected by the fluid channel 22, the culture medium can flow from one end of the culture unit 20 to the other end along the arrangement direction of the base holes 21. During the flow of the culture medium, the starting end of the culture medium flow is called the upstream end of the culture unit 20, and the other opposite end of the culture unit 20 is called the downstream end. The upstream end of each culture unit 20 is equipped with an upstream liquid storage pool 23, and the downstream end is equipped with a downstream liquid storage pool 24. The upstream liquid storage pool 23 and the most upstream base hole 21, and the downstream liquid storage pool 24 and the most downstream base hole 21 are also connected by the fluid channel 22. When the culture medium needs to be replaced, new culture medium is injected into the upstream liquid reservoir 23, and the plate body 10 is tilted in such a way that the upstream liquid reservoir 23 is higher than the downstream liquid reservoir 24, so that the original culture medium in the base hole 21 in the culture unit 20 flows to the downstream liquid reservoir 24 along the fluid channel 22, and the new culture medium in the upstream liquid reservoir 23 flows to each base hole 21 along the fluid channel 22. By making the new culture medium in the upstream liquid reservoir 23 continue to flow to the base hole 21 along the fluid channel 22, the original culture medium in the base hole 21 can be basically replaced eventually. In addition, by establishing a micro-power system (not shown) composed of a micro pump and a hose between the upstream liquid reservoir 23 and the downstream liquid reservoir 24, the culture medium flowing from the base hole 21 into the downstream liquid reservoir 24 can be continuously pumped into the upstream liquid reservoir 23, and the pumped culture medium continues to flow into each base hole 21 of the culture unit 20, so that the culture medium circulates between the base holes 21, so that the cells grow in the flowing culture medium.

[0034] In some preferred structures, a liquid inlet interface 231 is configured at a position of the upstream end surface of the plate body 10 in the length direction corresponding to the upstream liquid storage tank 23, and a liquid outlet interface 241 is configured at a position of the downstream end surface of the plate body 10 in the length direction corresponding to the downstream liquid storage tank 24. Both ends of the hose of the micro-power system are respectively connected to the liquid inlet interface 231 and the liquid outlet interface 241, so that the culture medium in the downstream liquid storage tank 24 is pumped into the upstream liquid storage tank 23 by means of the micro pump installed on the hose.

[0035] In some preferred structures, such as Figure 6 As shown, the bottom of each base hole 21 is lower than the highest area of ​​the bottom of the fluid channel 22 on both sides of the base hole 21, so that when the plate body 10 is in a horizontal state, the culture medium can always be retained in the base hole 21, so that cells can be cultured when the plate body 10 is in a horizontal state.

[0036] A key point of the present invention is that a downstream pore wall is configured between the fluid channel 22 downstream of the base pore 21 and the base pore 21, and a downstream pore wall is configured between the fluid channel 22 upstream of the base pore 21 and the base pore 21, and a plurality of downstream slits penetrating the downstream pore wall are processed on the downstream pore wall, and a plurality of upstream slits penetrating the upstream pore wall are processed on the upstream pore wall. The downstream slits and the upstream slits are used to allow the culture medium to pass through and restrict the passage of cells. In this way, during the process of replacing the culture medium or circulating the culture medium using a micro-power system, the cells will be blocked by the downstream pore wall to prevent the cells from being transferred to the downstream base pore 21 or even flowing into the downstream liquid storage pool 24 with the flow of the culture medium. In addition, the upstream pore wall and the downstream pore wall cooperate to restrict the cells to be cultured in the base pore 21 and prevent them from being transferred to the fluid channel 22 between the base pores 21.

[0037] The advantage of the above-mentioned structure for limiting the undesirable transfer of cells is that the above-mentioned structure is easier to configure and mold. Specifically, when the relevant structure (such as the fluid channel 22 and the base hole 21) has only a millimeter-level size, it is easier to process the above-mentioned structure than to assemble a filter screen, and the effect is better. The above-mentioned structure can be processed in the following manner: the fluid channel 22 and the base hole 21 can be integrally formed with the plate body 10 by injection molding and can be engraved by a laser engraving machine. Whether the fluid channel 22 and the base hole 21 are formed by injection molding or laser engraving, after the fluid channel 22 and the base hole 21 are formed, the solid hole wall is retained between the base hole 21 and the fluid channels 22 on both sides, and then the laser engraving machine is used again to cut the solid hole wall from the top of the plate body 10, thereby cutting out the upstream gap and the downstream gap. It can be seen that by configuring the structure for limiting cell transfer into the above-mentioned structural form, it can be processed by a laser engraving machine that can achieve millimeter and micrometer-level engraving.

[0038] In some preferred structures, such as Figure 5and Figure 6 , and combined with Figure 2 As shown, a downstream flow-increasing cavity extending in the circumferential direction is arranged between the downstream fluid channel 22 of the base hole 21 and the downstream hole wall, the downstream fluid channel 22 of the base hole 21 passes through to the downstream flow-increasing cavity, and the width of the downstream flow-increasing cavity is greater than the width of the port of the downstream fluid channel 22 of the base hole 21 so that the width of the downstream hole wall is greater than the width of the port of the downstream fluid channel 22 of the base hole 21; an upstream flow-increasing cavity extending in the circumferential direction is arranged between the upstream fluid channel 22 of the base hole 21 and the upstream hole wall, the upstream fluid channel 22 of the base hole 21 passes through to the upstream flow-increasing cavity, and the width of the upstream flow-increasing cavity is greater than the width of the port of the upstream fluid channel 22 of the base hole 21 so that the width of the upstream hole wall is greater than the width of the port of the upstream fluid channel 22 of the base hole 21. The circumferential extension scale of the upstream flow-increasing cavity and the downstream flow-increasing cavity of each base hole 21 is preferably 90° to 180° when converted by occupying the central angle of a circle, so that the upstream gap on the upstream hole wall and the downstream gap on the downstream hole wall can preferably cover nearly 90° to 180° when converted by occupying the central angle of a circle. The advantage of such a setting is that the flow cross section of the gap on the hole wall can be significantly increased, so that the flow cross section of the gap is basically equal to the flow cross section of the fluid channel 22, or even larger than the flow structure of the fluid channel 22, so that when the culture medium passes through the gap of the hole wall, the resistance of the hole wall to the culture medium is not large, which improves the smoothness of the flow of the culture medium. The above-mentioned flow-increasing cavity can also be engraved and formed by a laser engraving machine.

[0039] In some more preferred structures, all the base holes 21 of each culture unit 20 are located in the same plane; the bottom of the upstream flow-increasing cavity of each base hole 21 is lower than the bottom of the downstream flow-increasing cavity, and the bottom of the fluid channel 22 between two adjacent base holes 21 is an inclined surface, and the two ends of the inclined surface are respectively connected to the bottom of the downstream flow-increasing cavity and the bottom of the upstream flow-increasing cavity; wherein: the bottom of the upstream gap is not higher than the bottom of the corresponding upstream flow-increasing cavity, and the bottom of the downstream gap is not lower than the bottom of the corresponding downstream flow-increasing cavity. The advantage of such a configuration is that during the replacement of the culture medium or the circulation of the culture medium by the micro-power system, as the culture medium passes through the base hole 21 along the fluid channel 22, since the bottom of the downstream flow-increasing cavity and the downstream gap on the downstream side of the base hole 21 are higher than the bottom of the upstream flow-increasing cavity and the upstream gap on the upstream side of the base hole 21, the lower layer of the culture medium in the upstream fluid channel 22 will flow into the lower part of the base hole 21 after passing through the lower part of the upstream gap. Subsequently, this part of the culture medium encounters resistance and moves upward and then flows out through the downstream gap, so that the culture medium forms a wave-like flow, thereby accelerating the replacement of the culture medium in the lower part of the base hole 21, which is beneficial to improving the replacement efficiency and circulation efficiency of the culture medium.

[0040] It should be noted that when the plate body 10 is in a horizontal state, the liquid level of the culture medium in the base hole 21 is limited by the bottom of the downstream gap.

[0041] The base hole 21 includes a circular hole section at the top and a conical hole section at the bottom; the bottom of the downstream gap is located at the circular hole section, and the bottom of the upstream gap is located at the conical hole section. The advantages of such a configuration are: on the one hand, when it is necessary to remove cells from the base hole 21 by suction from the top of the plate body 10, the lower part of the base hole 21 is configured as a conical hole end, which makes it easier to suck and remove cells; on the other hand, the conical hole section is conducive to the flow of culture medium, thereby improving the replacement efficiency of the culture medium at the bottom of the base hole 21.

[0042] In some preferred structures, such as Figure 7 Combined with Figure 1 As shown, an upstream notch 233 extending downward from the top is formed on the upstream partition 232 between each adjacent upstream liquid storage pool 23, and a downstream notch 243 extending downward from the top is formed on the downstream partition 242 between each adjacent downstream liquid storage pool 24. The notch and the downstream notch 243 can be formed as follows: Figure 7 The blocking member 25 shown is blocked in a detachable manner. When in the blocking state, the two adjacent liquid storage pools are isolated from each other, and when the blocking member 25 is removed, the two adjacent liquid storage pools are interconnected. The purpose of such a configuration is that the culture medium in the two adjacent culture units 20 can flow to each other through the gap on the partition.

Claims

1. A cell culture device, characterized in that: include: plate body; A culture unit, comprising a plurality of culture units, which are arranged on the plate body at intervals along the width direction of the plate body, each culture unit comprising a plurality of base holes for culturing cells arranged at intervals along the length direction of the plate body and a fluid channel between each two adjacent base holes, and according to the flow direction of the culture medium along the fluid channel, the two sections of the fluid channel corresponding to the two sides of each base hole in the radial direction are called the fluid channel upstream of the base hole and the fluid channel downstream of the base hole; wherein: A downstream hole wall is configured between at least the fluid channel downstream of the basic hole and the basic hole, and a plurality of downstream gaps that allow the medium to pass through but restrict the cells to pass through are processed on the downstream hole wall by cutting.

2. The cell culture device according to claim 1, characterized in that: An upstream hole wall is arranged between the fluid channel upstream of the basic hole and the basic hole, and a plurality of upstream slits are processed on the upstream hole wall by cutting to allow the medium to pass through but restrict the cells to pass through.

3. The cell culture device according to claim 2, characterized in that: A downstream flow-increasing cavity extending in the circumferential direction is arranged between the downstream fluid channel of the base hole and the downstream hole wall, the downstream fluid channel of the base hole passes through the downstream flow-increasing cavity, the width of the downstream flow-increasing cavity is greater than the width of the port of the downstream fluid channel of the base hole so that the width of the downstream hole wall is greater than the width of the port of the downstream fluid channel of the base hole; An upstream flow-increasing cavity extending in the circumferential direction is arranged between the fluid channel upstream of the base hole and the upstream hole wall, the fluid channel upstream of the base hole passes through the upstream flow-increasing cavity, the width of the upstream flow-increasing cavity is greater than the width of the port of the fluid channel upstream of the base hole so that the width of the upstream hole wall is greater than the width of the port of the fluid channel upstream of the base hole.

4. The cell culture device according to claim 3, characterized in that: All the base holes of each culture unit are located in the same plane; the bottom of the upstream flow-increasing cavity of each base hole is lower than the bottom of the downstream flow-increasing cavity, and the bottom of the fluid channel between two adjacent base holes is an inclined surface, and the two ends of the inclined surface are respectively connected to the bottom of the downstream flow-increasing cavity and the bottom of the upstream flow-increasing cavity; wherein: the bottom of the upstream gap is not higher than the bottom of the corresponding upstream flow-increasing cavity, and the bottom of the downstream gap is not lower than the bottom of the corresponding downstream flow-increasing cavity.

5. The cell culture device according to claim 4, characterized in that: The base hole comprises a circular hole section located at an upper portion and a tapered hole section located at a lower portion.

6. The cell culture device according to claim 5, characterized in that: The bottom of the downstream gap is located in the circular hole section, and the bottom of the upstream gap is located in the tapered hole section.

7. The cell culture device according to claim 1, characterized in that: The culture unit further includes an upstream liquid storage tank located upstream of the most upstream base hole and a downstream liquid storage tank located downstream of the most downstream base hole.

8. The cell culture device according to claim 7, characterized in that: The two upstream liquid storage tanks and the two downstream liquid storage tanks of two adjacent culture units are separated by partitions; wherein: A notch extending downward from the top is formed on each of the partitions, and a blocking component is detachably mounted on the notch.

9. The cell culture device according to claim 7, characterized in that: Two end surfaces in the length direction of the plate body are provided with liquid inlet interfaces corresponding to the plurality of upstream liquid storage tanks one by one and liquid outlet interfaces corresponding to the plurality of downstream liquid storage tanks one by one.

10. The cell culture device according to claim 2, characterized in that: The upstream hole wall and the upstream slit on the upstream hole wall, the downstream hole wall and the downstream slit on the downstream hole wall are all obtained by laser cutting.

Images