Cell culture device

Abstract

The present invention relates to the field of microbial culture technology. The present invention discloses a cell culture device, comprising: a plate body; a plurality of culture units, wherein the plurality of culture units are arranged on the plate body at intervals along the width direction of the plate body, and each culture unit comprises 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 adjacent two base holes. According to the flow direction of the culture medium along the fluid channel, the two sections of fluid channels corresponding to the two sides in the radial direction of each base hole are called the fluid channel upstream of the base hole and the fluid channel downstream of the base hole; a downstream hole wall is configured at least between the fluid channel downstream of the base hole and the base hole, and a plurality of downstream gaps that allow the culture medium to pass through but restrict the passage of cells are processed on the downstream hole wall in a cutting manner.

Description

Technical Field

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

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

[0003] The above-mentioned well plates in the prior art have the following drawbacks for culturing cells: the wells are independent of each other. When the culture medium needs to be replaced, a device with multiple needles arranged in a consistent pattern must be used to simultaneously aspirate and inject the culture medium into each well. Otherwise, a single needle must be used to aspirate and inject the culture medium into each well in sequence. Therefore, the well plates with the above-mentioned structure require specialized and expensive equipment to complete the culture medium replacement, and the well plates must be placed in the environment where the equipment is located. In addition, in some cases, for example, if the cells are small in size, when the culture medium is replaced by aspiration using a needle, the cells may be simultaneously sucked into the needle, resulting in the undesirable removal of the cells.

[0004] To overcome these drawbacks, a well plate has emerged in the prior art for facilitating culture medium replacement (or transfer). Specifically, this well plate improves upon conventional well plates by connecting each matrix-arranged column (row) of wells via a fluid channel. Specifically, a fluid channel connecting two adjacent wells in each column (row) is machined between them. This allows medium removal by aspirating from the rear of each column (row) or by tilting the well plate to direct medium from each well toward the rear. Furthermore, medium replacement can be achieved by injecting medium into the wells at the head, gradually dispensing it into each well.

[0005] However, the improved orifice plates of the prior art still suffer from the following drawbacks during use: during the process of removing culture medium through the fluid channels and injecting culture medium into the basal wells, some cells in the basal wells may be transferred with the culture medium to other basal wells, or even flow out of the plate with the culture medium, resulting in the undesirable removal of cells. Although technicians have attempted to prevent cell transfer by inserting filters into the basal wells, these measures have ultimately proved difficult to implement because the basal wells are generally sized in the millimeter or even micrometer range, making filters suitable for these sizes difficult to prepare and 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:

[0008] A cell culture device comprising:

[0009] plate body;

[0010] A culture unit, comprising a plurality of culture units, each of which is spaced apart along the width direction of the plate body on the plate body, each culture unit comprising a plurality of base holes for culturing cells spaced apart along the length direction of the plate body and a fluid channel between each adjacent two base holes, wherein, according to the flow direction of the culture medium along the fluid channel, the two fluid channels corresponding to the two sides of each base hole in the radial direction are referred to as the fluid channel upstream of the base hole and the fluid channel downstream of the base hole; wherein:

[0011] A downstream pore wall is configured at least between the fluid channel downstream of the basic pore and the basic pore, and a plurality of downstream slits that allow the medium to pass through but restrict the cells to pass through are processed on the downstream pore wall by cutting.

[0012] Preferably, an upstream pore wall is configured between the fluid channel upstream of the basic pore and the basic pore, and a plurality of upstream slits that allow the medium to pass through but restrict the passage of cells are processed on the upstream pore wall by cutting.

[0013] Preferably,

[0014] 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;

[0015] 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.

[0016] Preferably, all the basal holes of each culture unit are located in the same plane; the bottom of the upstream flow-increasing cavity of each basal hole is lower than the bottom of the downstream flow-increasing cavity, and the bottom of the fluid channel between two adjacent basal 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 slit is not higher than the bottom of the corresponding upstream flow-increasing cavity, and the bottom of the downstream slit is not lower than the bottom of the corresponding downstream flow-increasing cavity.

[0017] Preferably, the base hole comprises a circular hole section located at the upper portion and a tapered hole section located at the lower portion.

[0018] 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.

[0019] 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.

[0020] 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:

[0021] A notch extending downward from the top is formed on each of the partitions, and a blocking component is detachably mounted on the notch.

[0022] 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.

[0023] 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.

[0024] Compared with the prior art, the cell culture device provided by the present invention has the following beneficial effects:

[0025] The present invention configures a pore wall with a gap between the fluid channel and the base pore, so that during the replacement of culture medium or the recycling of culture medium, the pore wall allows the culture channel to pass while restricting the passage of cells, thereby avoiding undesirable cell migration.

[0026] This disclosure is an overview of various implementations or examples of the technology, and is not a comprehensive disclosure of the full scope or all features of the disclosed technology. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In the drawings, which are not necessarily drawn to scale, the same reference 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 generally illustrate various embodiments by way of example and not limitation, and together with the description and claims, serve to illustrate the embodiments of the invention. Where 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 of the embodiments of the present apparatus or method.

[0028] Figure 1This is a schematic diagram of the three-dimensional structure of the cell culture device provided by the present invention.

[0029] Figure 2 for Figure 1 An enlarged view of detail A.

[0030] Figure 3 This is a top view of the cell culture device provided by the present invention.

[0031] Figure 4 for Figure 3 An enlarged view of detail B.

[0032] Figure 5 This is a three-dimensional cross-sectional view of the cell culture device provided by the present invention.

[0033] Figure 6 for Figure 5 An enlarged view of a portion C.

[0034] Figure 7 This is a schematic structural diagram of the blocking component in the cell culture device provided by the present invention.

[0035] Reference numerals:

[0036] 10-plate body; 20-culture unit; 21-basal 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

[0037] Unless otherwise defined, the technical or scientific terms used in the present invention shall have the usual meanings understood by persons of ordinary skill in the field to which the present invention belongs. The words "first", "second" and similar terms used in the present invention do not indicate any order, quantity or importance, but are only used to distinguish different components. Words such as "include" or "comprise" mean that the elements or objects preceding the word include the elements or objects listed after the word and their equivalents, without excluding other elements or objects. Words such as "connect" or "connected" 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 object being described changes, the relative positional relationship may also change accordingly.

[0038] In order to keep the following description of the embodiments of the present invention clear and concise, detailed descriptions of known functions and known components are omitted.

[0039] like Figures 1 to 7 As shown, the present invention discloses a cell culture device for culturing a large number of cells simultaneously. The device includes a plate body 10 and a plurality of culture units 20 arranged on the upper surface of the plate body 10 .

[0040] The plate body 10 can be made of a biocompatible material, for example, it can be made of a biocompatible high molecular polymer by injection molding, or it can be made of a biocompatible alloy material, for example, titanium alloy, or it can be composite molded of titanium alloy and high molecular polymer. Preferably, the plate body 10 is made of titanium alloy or a composite with a titanium alloy on the upper part and a high molecular polymer on the lower part.

[0041] like Figures 1 to 4 As shown, the culture units 20 include a plurality of culture units 20, which are arranged at intervals along the width of the plate body 10 and disposed on the upper portion of the plate body 10. Each culture unit 20 includes a plurality of base holes 21 spaced apart along the length of the plate body 10, and a fluid channel 22 between each two base holes 21. The fluid channel 22 interconnects the base holes 21 to allow fluid flow. Each base hole 21 is used to culture cells. Therefore, each base hole 21 can be implanted with cells, and the cells can be cultured by injecting culture medium into the base hole 21.

[0042] Because each adjacent pair of basal pores 21 is connected by a fluid channel 22, culture medium can flow from one end of the culture unit 20 to the other along the arrangement of the basal pores 21. During this flow, the culture medium passes through the basal pores 21. Based on the direction of culture flow, the starting point of culture flow is referred to as the upstream end of the culture unit 20, while the opposite end of the culture unit 20 is referred to as the downstream end. Each culture unit 20 is equipped with an upstream liquid reservoir 23 at its upstream end and a downstream liquid reservoir 24 at its downstream end. Fluid channels 22 also connect the upstream liquid reservoir 23 to the upstreammost basal pore 21, and the downstream liquid reservoir 24 to the downstreammost basal pore 21. When culture medium replacement is required, new culture medium is injected into the upstream reservoir 23, and the plate 10 is tilted so that the upstream reservoir 23 is higher than the downstream reservoir 24. As a result, the original culture medium in the basal wells 21 of the culture unit 20 flows along the fluid channel 22 to the downstream reservoir 24, and the new culture medium in the upstream reservoir 23 flows along the fluid channel 22 to each basal well 21. By continuously allowing the new culture medium in the upstream reservoir 23 to flow along the fluid channel 22 to the basal wells 21, the original culture medium in the basal wells 21 can eventually be substantially replaced. Furthermore, by establishing a micro-power system (not shown) consisting of a micropump and a hose between the upstream reservoir 23 and the downstream reservoir 24, the culture medium flowing from the basal well 21 into the downstream reservoir 24 can be continuously pumped into the upstream reservoir 23. The pumped culture medium then flows into each basal well 21 of the culture unit 20, thereby circulating the culture medium between the basal wells 21 and allowing cells to grow in the flowing culture medium.

[0043] In some preferred structures, a liquid inlet interface 231 is configured at a position corresponding to the upstream end face of the plate body 10 in the longitudinal direction and the upstream liquid storage tank 23, and a liquid outlet interface 241 is configured at a position corresponding to the downstream end face of the plate body 10 in the longitudinal direction and the downstream liquid storage tank 24. The two 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.

[0044] 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. Therefore, when the plate body 10 is in a horizontal state, culture medium can always be retained in the base hole 21. In this way, cells can be cultured when the plate body 10 is in a horizontal state.

[0045] 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 confine 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.

[0046] The advantage of the aforementioned structure for restricting unwanted cell migration is that it is easier to configure and mold. Specifically, when the relevant structures (such as the fluid channel 22 and the base hole 21) are only millimeter-scale in size, it is easier to manufacture the aforementioned structure compared to assembling a filter, and the effect is better. The aforementioned structure can be manufactured as follows: the fluid channel 22 and the base hole 21 can be integrally molded with the plate 10 by injection molding, or they can be engraved using a laser engraving machine. Whether the fluid channel 22 and the base hole 21 are injection molded or laser engraved, after the fluid channel 22 and the base hole 21 are formed, a solid pore wall is retained between the base hole 21 and the fluid channels 22 on either side. Then, a laser engraving machine is used to cut the solid pore wall from above the plate 10, thereby cutting the upstream and downstream slits. Thus, by configuring the structure for restricting cell migration into the aforementioned structural form, it can be processed using a laser engraving machine capable of millimeter- and micron-scale engraving.

[0047] In some preferred structures, such as Figure 5 and Figure 6 , and combined with Figure 2As 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 of the upstream flow-increasing cavity and the downstream flow-increasing cavity of each base hole 21 is preferably 90° to 180° when converted to the central angle of the circle, so that the upstream gap on the upstream hole wall and the downstream gap on the downstream hole wall can be preferably distributed close to 90° to 180° when converted to the central angle of the 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. Therefore, when the culture medium passes through the gap in the hole wall, the hole wall has little resistance to the culture medium, which improves the smoothness of the flow of the culture medium. The above-mentioned flow-increasing cavity can also be engraved using a laser engraving machine.

[0048] In some more preferred structures, all the basic holes 21 of each culture unit 20 are located in the same plane; the bottom of the upstream flow-increasing cavity of each basic hole 21 is lower than the bottom of the downstream flow-increasing cavity, and the bottom of the fluid channel 22 between two adjacent basic holes 21 is a slope, and the two ends of the slope 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 using 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.

[0049] 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.

[0050] The base hole 21 comprises an upper circular section and a lower tapered section. The bottom of the downstream slit is located in the circular section, while the bottom of the upstream slit is located in the tapered section. This configuration offers the following advantages: When cells need to be removed from the base hole 21 by suction from above the plate 10, the tapered end of the lower portion of the base hole 21 facilitates suction and removal. Furthermore, the tapered section facilitates the flow of culture medium, thereby improving the efficiency of culture medium replacement in the lower portion of the base hole 21.

[0051] 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 tank 23, and a downstream notch 243 extending downward from the top is formed on the downstream partition 242 between each adjacent downstream liquid storage tank 24. The notch and the downstream notch 243 can be formed as follows Figure 7 The blocking member 25 is shown as a detachable blocking member. When in the blocking state, the two adjacent liquid reservoirs are isolated from each other. When the blocking member 25 is removed, the two adjacent liquid reservoirs are interconnected. This configuration allows the culture medium in the two adjacent culture units 20 to flow through the gaps in the partition.

Claims

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

2. The cell culture device according to claim 1, characterized in that All the basal holes of each culture unit are located in the same plane; the bottom of the upstream flow-increasing cavity of each basal hole is lower than the bottom of the downstream flow-increasing cavity; the bottom of the fluid channel between two adjacent basal holes is a slope, and the two ends of the slope 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 slit is not higher than the bottom of the corresponding upstream flow-increasing cavity, and the bottom of the downstream slit is not lower than the bottom of the corresponding downstream flow-increasing cavity.

3. The cell culture device according to claim 2, characterized in that The basic hole includes a circular hole section located at the upper portion and a tapered hole section located at the lower portion.

4. The cell culture device according to claim 3, 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.

5. The cell culture device according to claim 1, wherein 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.

6. The cell culture device according to claim 5, 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.

7. The cell culture device according to claim 5, characterized in that Liquid inlet interfaces corresponding to the plurality of upstream liquid storage tanks and liquid outlet interfaces corresponding to the plurality of downstream liquid storage tanks are respectively opened on the two end surfaces in the length direction of the plate body.

8. The cell culture device according to claim 1, wherein 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.

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