Deep-hole cell culture plate

Abstract

The utility model provides a deep hole cell culture plate which comprises a deep hole plate provided with a plate body, the plate body is provided with a plurality of deep holes and one or more groups of air holes with filter membranes, the upper surface of the plate body is provided with a membrane layer covering the orifices of all the deep holes in an adhesive mode, and the membrane layer is provided with a plurality of air holes. One or more groups of air holes with filter membranes are formed in the membrane layer; the plate groove is provided with an inner container and a shell arranged outside the inner container in a sleeving mode, and a cavity which is provided with an opening in the upper portion and used for containing the deep hole plate is formed in the inner container; the plate cover is arranged above the plate groove and used for covering an opening of the plate groove, and one or more sets of air holes with filter membranes are further formed in the upper surface of the plate cover. The ready-to-use deep-hole cell culture plate provided by the utility model is low in cost and easy to operate, can be used for high-quality culture and expression of cells in biological products, and can be used for carrying out instant detection on the cells and expression products thereof.

Description

Technical Field

[0001] This invention belongs to the field of cell culture and expression product detection, and specifically relates to a deep-well cell culture plate. Background Technology

[0002] Biopharmaceuticals, including vaccines, bacterial vaccines, antibiotics, bioactive substances, and antibodies, are intermediate products or secretions of metabolism within engineered cells. Cell culture plays a crucial role in the efficient production of these biopharmaceuticals, and a good cell culture state depends on numerous conditions, such as the culture environment, culture medium, and culture equipment. Among these, the culture equipment plays a vital role in the cell culture state, and the quality of the cells has a decisive impact on the quality of subsequent products or experiments.

[0003] Currently, the main biological equipment used for cell culture includes: 1) large culture jars or culture shake bags: mainly used for large-volume, high-volume biopharmaceutical production, such as the culture of monoclonal antibodies; 2) culture flasks: mainly used for research and development, pilot production, etc.; 3) disposable 96-well culture plates: mainly used for research and development screening, biological sample activity detection, etc.

[0004] Currently, there is no ready-to-use cell culture tool that can be directly used for the rapid and high-quality acquisition and immediate testing of cell products. Therefore, it is particularly important to develop a ready-to-use consumable for cell culture in biopharmaceuticals, which must ensure the cell culture status while also enabling immediate testing to quickly obtain experimental results and thus ensure the quality control of biopharmaceuticals. Utility Model Content

[0005] To address the problem that existing cell culture tools cannot simultaneously meet the requirements of high-quality cell culture and real-time detection, this invention provides a low-cost, easy-to-use, ready-to-use deep-well cell culture plate for high-quality cell culture and expression in biopharmaceuticals, and for real-time detection of cells and their expression products.

[0006] The technical solution adopted in this utility model is: a deep-well cell culture plate, comprising:

[0007] A deep-hole plate has a plate body on which multiple deep holes and one or more sets of air vents with filter membranes are formed. The upper surface of the plate body is adhesively provided with a membrane layer covering all the openings of the deep holes. The membrane layer has one or more sets of air vents with filter membranes.

[0008] The plate groove has an inner liner and an outer shell fitted outside the inner liner, the inner liner forming a cavity with an opening at the top for accommodating the deep hole plate;

[0009] A cover is disposed above the plate groove to cover the opening of the plate groove, and the upper surface of the cover is provided with one or more sets of air vents with filter membranes.

[0010] In some embodiments, the cavity of the plate groove also has a cold accumulator.

[0011] In some embodiments, the inner container of the plate groove is made of stainless steel.

[0012] In some embodiments, a thermal insulation layer is arranged between the inner container and the outer shell of the plate groove, and the thermal insulation layer has a thermal insulation material.

[0013] In some embodiments, the inner surface of the plate cover is provided with a first clamping part which is detachably assembled to a second clamping part arranged on the outer surface of the plate groove.

[0014] In some embodiments, the edge of the plate body extends outwardly to form a support edge, the deep-well plate is erected on the opening of the plate groove through the support edge, and the plate cover is covered on the opening of the plate groove through the deep-well plate.

[0015] In some embodiments, the opening of the plate groove is circumferentially provided with a first upper sealing member which is tightly attached to a first lower sealing member circumferentially arranged on the lower end surface of the plate body, and the cover of the plate cover is circumferentially provided with a second upper sealing member which is tightly attached to a second lower sealing member circumferentially arranged on the upper end surface of the plate body.

[0016] In some embodiments, the plate body is arrayed with 12, 24, 48 or 96 deep wells.

[0017] In some embodiments, the bottom of the deep well is conical.

[0018] In some embodiments, the orifice of the deep well is rectangular or circular.

[0019] The utility model also provides a detection equipment, including detection equipment and deep well cell culture plate, the detection equipment is equipped with installation base plate, the deep well cell culture plate is placed in the detection equipment through the installation base plate.

[0020] The utility model provides a ready-to-use deep well cell culture plate which effectively solves the problem that high-quality cell culture and instant detection cannot be considered together, has the advantages of low cost and easy operation. The cell culture plate places the deep well plate convenient for detection in the closed space enclosed by the plate groove and the plate cover, and provides a good growth environment for the cells in the deep well through the air permeable hole with the filter membrane, ensures the stability of the cells in the culture process and the isolation from the external environment. At the same time, the deep well plate can be combined with the detection equipment to meet the demand of instant detection, thereby forming an efficient cell culture and detection system, effectively guaranteeing the quality control of biological products. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 For the deep hole cell culture plate in the embodiment of the utility model, wherein, 10 - plate cover, 101 - air hole, 20 - deep hole plate, 201 - plate body, 202 - deep hole, 30 - plate groove, 301 - inner container, 302 - outer shell. DETAILED DESCRIPTION

[0022] The embodiments of the utility model are explained below through specific specific embodiments, and other advantages and effects of the utility model can be easily understood by those skilled in the art from the disclosure of the specification. The utility model can also be implemented or applied through other different specific embodiments, and each detail in the specification can be modified or changed based on different viewpoints and applications without departing from the spirit of the utility model. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict.

[0023] In order to solve the problem that the cell culture tool in the prior art cannot simultaneously meet the high-quality culture and instant detection of cells, the embodiment of the utility model provides a low-cost, easy-to-operate instant deep hole cell culture plate for high-quality culture and expression of cells in biological products, and instant detection of expression products.

[0024] Please refer to Figure 1 , the deep hole cell culture plate includes deep hole plate 20, plate groove 30, plate cover 10. The deep hole plate 20 has plate body 201, a plurality of deep holes 202 and one or more groups of air holes with filter membrane (not shown in the figure) are formed on the plate body 201, and a film layer (not shown in the figure) covering all deep hole 202 orifices is arranged on the upper surface of the plate body 201 in a pasting manner, and the film layer is provided with one or more groups of air holes with filter membrane (not shown in the figure). The plate groove 30 has inner container 301 and outer shell 302 sleeved on the outer part of the inner container 301, and the inner container 301 forms a cavity for accommodating the deep hole plate 20 with an upper opening. The plate cover 10 is arranged above the plate groove 30 for covering the opening of the plate groove 30, and the upper surface of the plate cover 10 is also provided with one or more groups of air holes 101 with filter membrane.

[0025] In Figure 1In the illustrated embodiment, the deep-well plate 20 is placed inside the cavity of the plate groove 30, and then the plate cover 10 is placed on top of the plate groove 30 to assemble a complete deep-well cell culture plate with a closed space. The deep-well plate 20 has a plate body 201, on which 8×12 identical deep wells 202 are formed, and one or more sets of vent holes with filter membranes distributed between the deep wells 202 (not shown in the figure). The deep-well plate 20 can be integrally molded from polypropylene. The bottom of the deep well 202 is V-shaped, and the opening is circular. The deep wells 202 are used to contain cell culture medium and cultured cells. The depth of the deep wells 202 is greater than the pore depth of a typical 96-well plate, thereby providing a safe and stable culture environment for cells and avoiding external contamination or cross-contamination between wells. When transporting the deep-well plate 20 containing cells and their expression products, a coolant (such as liquid nitrogen) can be added to the cavity of the plate groove 30. One or more sets of vents with filter membranes on the plate body 201 allow the gas generated by the vaporization of liquid nitrogen to escape promptly, thus preventing the plate groove 30 from exploding due to gas expansion in the enclosed space, and also preventing contamination between upper and lower layers. To improve the airtightness and permeability of the deep-well cell culture plate, a membrane layer (not shown in the figure) can be adhesively applied to the upper surface of the plate body 201 to cover and seal the openings of all deep wells 202. The membrane layer minimizes the evaporation of liquid within the deep wells 202 and protects the liquid from contamination and splashing. The membrane layer can be made of materials such as vinyl plastic, polypropylene, or polyester acrylate. Simultaneously, one or more sets of vents with filter membranes (not shown in the figure) are also provided on the membrane layer. These vents with filter membranes not only improve the permeability of the deep-well cell culture plate but also prevent external pathogens from contaminating the cells within the wells. The plate groove 30 has an inner liner 301 and an outer shell 302 fitted over the inner liner 301. The inner liner 301 forms a cavity with an opening at the top for accommodating the deep-well plate 20. The deep-well plate 20 is placed in the inner liner 301 of the plate groove 30, and then the plate cover 10, located above the plate groove 30, closes the opening of the plate groove 30. Through the combined effects of the inner liner 301, the outer shell 302, and the plate cover 10, the cells inside the deep-well plate 20 are protected and well-insulated, preventing contamination or drastic temperature fluctuations. Furthermore, the upper surface of the plate cover 10 is provided with one or more sets of venting holes 101 with filter membranes. These venting holes 101 with filter membranes further improve the overall permeability of the deep-well cell culture plate and prevent external pathogens from contaminating the cells inside the wells.

[0026] right Figure 1 The illustrated embodiment underwent sterility testing, air permeability verification, and functional testing after cell cryopreservation and thawing. The test results demonstrate that this deep-well cell culture plate can provide a favorable growth environment for cells, ensuring cell stability during culture and isolation from the external environment.

[0027] Sterility test: According to the sterility test requirements, the cells were sampled for sterility test after being cultured in the deep-well cell culture plate for 2 days, and the result was negative.

[0028] Verification of air permeability: 2 / 3 of the liquid nitrogen was added to the inner container of the plate groove of the deep-well cell culture plate, and the three parts of the deep-well cell culture plate (deep-well plate, plate groove, and plate cover) were assembled and placed at room temperature. The liquid nitrogen in the inner container 301 was completely gasified, and the deep-well plate, plate groove, and plate cover were all intact.

[0029] Functional detection after cell cryopreservation and recovery: The deep-well cell culture plate with cells and complete assembly was cryopreserved (cells: HL-60 (P7+5+6), 1.5E7 / 0.5mL, viability: 90.48%; cryopreservation solution: cell cryopreservation solution) and recovered, and MAT detection was performed. The detection results prove that after the cells HL-60 are cryopreserved and recovered by the deep-well cell culture plate, the linear range, intermediate precision, minimum detection limit (LOD 0.0125EU / mL), lower limit of quantification (LLOQ 0.025EU / mL), etc. of MAT are consistent with the results of HL-60 cells cultured normally.

[0030] In some embodiments, the number of deep-wells 202 arrayed on the plate body 201 can be 12, or 24, or 48, or 96. The opening mode of the plate body 201 is generally in accordance with the ANSI standard, and is compatible with various high-throughput workflow automation liquid handling and commonly used detection instruments. In some embodiments, the bottom of the deep-well 202 is U-shaped or V-shaped. For example, the U-shaped bottom helps to resuspend the cells, and the V-shaped bottom facilitates cell recovery. In some embodiments, the orifice of the deep-well 202 is rectangular or circular.

[0031] In some embodiments, to achieve better sealing effect, the inner surface of the plate cover 10 is provided with a first clamping part, which is detachably assembled to a second clamping part provided on the outer surface of the plate groove 30. Through the clamping mode, the plate cover 10 can be more tightly and firmly covered on the plate groove 30, preventing accidental falling.

[0032] In the process of transporting the deep-well plate 20 storing cells and their expression products, the heat preservation function of the plate groove 30 can also be improved to have better heat preservation effect. In some embodiments, the cavity of the plate groove 30 also has a cold accumulator, the inner container 301 of the plate groove 30 is made of stainless steel, and a heat preservation layer (not shown in the figure) is arranged between the inner container 301 and the outer shell 302, and the heat preservation layer has a heat preservation material such as foam. Specifically, liquid nitrogen can be selected as the cold accumulator and added to the inner container 301 made of stainless steel, and then the deep-well plate 20 is placed in the inner container 301, and the plate cover 10 is closed. Under the action of liquid nitrogen and the heat preservation layer, the deep-well cell culture plate has better heat preservation effect, which facilitates the transportation of the deep-well cell culture plate and prevents the cells and their expression products from being affected by temperature fluctuations during transportation. In order to facilitate the taking of the deep-well plate 20 during detection, in some embodiments, the edge of the plate body 201 extends outward to form a support edge (not shown in the figure), and the outer edge of the support edge is flush with the outer edge of the upper end opening of the plate groove 30, so that the deep-well plate 20 can be erected on the opening of the plate groove 30, and the plate cover 10 is closed on the opening of the plate groove 30 through the deep-well plate 20. In this way, the deep-well plate 20 can be conveniently erected on the plate groove 30, and then the plate cover 10 is closed on the deep-well plate 20. In this embodiment, in order to achieve better sealing effect, the opening of the plate groove 30 is circumferentially provided with a first upper sealing member, and the first upper sealing member is tightly attached to a first lower sealing member circumferentially arranged on the lower end surface of the support edge; the cover of the plate cover 10 is circumferentially provided with a second upper sealing member, and the second upper sealing member is tightly attached to a second lower sealing member circumferentially arranged on the upper end surface of the support edge. The first sealing member formed by the cooperation between the support edge and the plate groove 30, and the second sealing member formed by the cooperation between the support plate and the plate cover 10 have better sealing effect between the deep-well plate 20, the plate groove 30 and the plate cover 10, preventing liquid from splashing or external pollution from entering.

[0033] The utility model embodiment further provides a kind of detection equipment, including detection equipment and the deep-well cell culture plate of described, the mounting substrate is equipped on the detection equipment, and the deep-well cell culture plate is placed in the detection equipment by the mounting substrate.In this embodiment, the deep-well cell culture plate storing cells and their expression products is placed in the detection equipment matched with it, and cell or its expression product is detected.

[0034] The above-described embodiments are merely preferred embodiments of the utility model, and do not limit the scope of the utility model, and various modifications and improvements to the technical solutions of the utility model made by those skilled in the art without departing from the design spirit of the utility model shall fall within the protection scope of the utility model.

Claims

1. A deep-well cell culture plate, characterized in that, include: A deep hole plate (20) has a plate body (201) on which a plurality of deep holes (202) and one or more sets of air vents with filter membranes are provided. The upper surface of the plate body (201) is provided with a membrane layer covering all the openings of the deep holes (202) in an adhesive manner. The membrane layer has one or more sets of air vents with filter membranes. The plate groove (30) has an inner liner (301) and an outer shell (302) sleeved on the outside of the inner liner (301), wherein the inner liner (301) forms a cavity with an opening at the top for accommodating the deep hole plate (20); A cover (10) is disposed above the plate groove (30) to cover the opening of the plate groove (30), and the upper surface of the cover (10) is also provided with one or more sets of air vents (101) with filter membrane.

2. The deep-well cell culture plate as described in claim 1, characterized in that, The cavity of the plate groove (30) also contains a cold storage agent.

3. The deep-well cell culture plate as described in claim 1, characterized in that, The inner liner (301) of the plate groove (30) is made of stainless steel.

4. The deep-well cell culture plate as described in claim 1, characterized in that, A heat insulation layer is also provided between the inner liner (301) and the outer shell (302) of the plate groove (30), and the heat insulation layer has heat insulation material.

5. The deep-well cell culture plate as described in claim 1, characterized in that, The inner surface of the cover (10) is provided with a first snap-fit ​​part, which is detachably assembled to a second snap-fit ​​part provided on the outer surface of the groove (30).

6. The deep-well cell culture plate as described in claim 1, characterized in that, The edge of the plate (201) extends outward to form a support edge. The deep hole plate (20) is mounted on the opening of the plate groove (30) through the support edge. The plate cover (10) is closed to the opening of the plate groove (30) through the deep hole plate (20).

7. The deep-well cell culture plate as described in claim 6, characterized in that, The opening of the plate groove (30) is provided with a first upper seal in the circumferential direction, and the first upper seal is closely fitted with a first lower seal in the circumferential direction on the lower end face of the support side; the cover edge of the plate cover (10) is provided with a second upper seal in the circumferential direction, and the second upper seal is closely fitted with a second lower seal in the circumferential direction on the upper end face of the support side.

8. The deep-well cell culture plate as described in claim 1, characterized in that, The plate (201) has 12, 24, 48, or 96 deep holes (202) arranged in an array.

9. The deep-well cell culture plate as described in claim 1, characterized in that, The bottom of the deep hole (202) is U-shaped or V-shaped; and / or the opening of the deep hole (202) is rectangular or circular.

10. A detection system, characterized in that, The device includes a detection device and a deep-well cell culture plate as described in any one of claims 1 to 9; the detection device is equipped with a mounting substrate, and the deep-well cell culture plate is placed in the detection device via the mounting substrate.

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