CN219470073U - In-vitro micro-tissue co-culture pore plate - Google Patents
In-vitro micro-tissue co-culture pore plate Download PDFInfo
- Publication number
- CN219470073U CN219470073U CN202320667080.0U CN202320667080U CN219470073U CN 219470073 U CN219470073 U CN 219470073U CN 202320667080 U CN202320667080 U CN 202320667080U CN 219470073 U CN219470073 U CN 219470073U
- Authority
- CN
- China
- Prior art keywords
- culture
- micro
- tissue
- liquid storage
- vitro
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000003501 co-culture Methods 0.000 title claims abstract description 31
- 239000011148 porous material Substances 0.000 title claims abstract description 29
- 238000000338 in vitro Methods 0.000 title claims abstract description 28
- 239000007788 liquid Substances 0.000 claims abstract description 37
- 238000003860 storage Methods 0.000 claims abstract description 29
- 230000005484 gravity Effects 0.000 abstract description 5
- 230000009471 action Effects 0.000 abstract description 4
- 238000010874 in vitro model Methods 0.000 abstract description 2
- 239000001963 growth medium Substances 0.000 description 13
- 210000001519 tissue Anatomy 0.000 description 12
- 238000003384 imaging method Methods 0.000 description 7
- 239000006285 cell suspension Substances 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 4
- 239000011159 matrix material Substances 0.000 description 3
- 230000003068 static effect Effects 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 2
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000007876 drug discovery Methods 0.000 description 2
- 210000002889 endothelial cell Anatomy 0.000 description 2
- 210000002744 extracellular matrix Anatomy 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000011664 nicotinic acid Substances 0.000 description 2
- 230000002792 vascular Effects 0.000 description 2
- 238000010146 3D printing Methods 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 108010085895 Laminin Proteins 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000036267 drug metabolism Effects 0.000 description 1
- 238000007877 drug screening Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000000017 hydrogel Substances 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 108010082117 matrigel Proteins 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 238000007639 printing Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The utility model discloses an in-vitro micro-tissue co-culture pore plate, which comprises a pore plate body, wherein at least one group of co-culture units are arranged on the pore plate body, and each group of culture units comprises: a liquid storage tank; a culture well; a sample adding hole; the liquid storage tank, the culture holes and the sample adding holes are arranged in pairs, each culture hole is provided with the sample adding hole, and the liquid storage tank is arranged at the outer side of each culture hole; each pair of the liquid storage tanks and each pair of the culture holes are communicated through a microfluidic channel. According to the utility model, different amounts of culture mediums are added into the liquid storage tanks at two sides of the culture hole, and the flow direction of the culture mediums is directionally driven by utilizing the gravity action, so that the multi-tissue combined dynamic co-culture in-vitro model is constructed.
Description
Technical Field
The utility model relates to the technical field of biological manufacturing, in particular to an in-vitro micro-tissue co-culture pore plate.
Background
At present, the traditional culture technology is mainly based on single-hole static culture, and a culture mode of high-throughput dynamic culture is fresh. The environment in the human body is the whole body of the cooperative work of the tissues and organs, and the in-vitro high-throughput construction of the multi-tissue co-culture system has great significance for exploring the cooperative growth among tissues, exploring the metabolism of the tissue medicines and the like. In addition, in tissue in-vitro culture, a 96-well plate is often used, and the aperture of the 96-well plate is larger, so that samples are randomly distributed in the culture holes, and the samples to be cultured are inconvenient to be effectively fixed in a specific visual field, so that shooting resource waste is caused.
The existing co-culture pore plate has a narrow application range, a single and static culture mode, and cannot construct a complex in-vitro culture microenvironment.
Disclosure of Invention
The utility model aims to provide an in-vitro micro-tissue co-culture pore plate, which solves the problems that the application range of the co-culture pore plate in the prior art is narrow, the culture mode is single and static, and a complex in-vitro culture micro-environment cannot be constructed.
The technical scheme adopted by the utility model is as follows:
an in vitro micro-tissue co-culture well plate, comprising a well plate body on which at least one group of co-culture units is arranged, each group of culture units comprising:
a liquid storage tank with one end being open and arranged on the upper surface of the pore plate body;
one end of the culture hole is open and is arranged on the upper surface of the pore plate body, and the other end of the culture hole is communicated with the liquid storage tank through a micro-flow channel;
the sample adding hole is opened at one end and is arranged on the upper surface of the pore plate body, and the other end of the sample adding hole is communicated with the culture hole through a micro-flow channel; and
The liquid storage tank, the culture holes and the sample adding holes are all arranged in pairs, each culture hole is provided with the sample adding hole, and the liquid storage tank is arranged at the outer side of each culture hole; each pair of the liquid storage tanks and each pair of the culture holes are communicated through a microfluidic channel.
Preferably, the culture well and the sample addition well are funnel-shaped with large top and small bottom, and the inclination angle of the funnel-shaped is 45-75 degrees.
Preferably, the funnel-shaped inclination angle is 70 degrees.
Preferably, the microfluidic channel comprises a first microfluidic channel, a second microfluidic channel and a third microfluidic channel, and the first microfluidic channel and the second microfluidic channel are oppositely arranged on two opposite sides of the third microfluidic channel.
Preferably, the connection parts of the first microfluidic channel, the third microfluidic channel, the second microfluidic channel and the third microfluidic channel are all connected with a sample adding microfluidic channel.
Preferably, the liquid storage tank is columnar.
Preferably, the pore diameter of the sample addition well is smaller than the pore diameter of the culture well.
Preferably, the center-to-center distance between the reservoir and the culture well is 8 to 10mm.
The beneficial effects of the utility model at least comprise:
1. the method is simple and convenient to operate, and is suitable for constructing in vitro co-culture models of various different tissues.
2. The compatibility is high. Based on the current commercial culture pore plate structure, the device is compatible with various high-flux imaging devices, pipetting workstations and the like.
3. Complex microenvironments may be built. The vascular microenvironment may be constructed by adding a cell matrix, cell suspension, etc., such as a mixed cell suspension of endothelial cells and matrix, to the loading microwells.
4. Constructing a more bionic multi-organ co-culture network. By culturing the micro-tissues at different locations in different culture chambers, a bionic in vitro system is constructed. For example: the heart-lung-liver-kidney circulatory system is formed by respectively inoculating heart micro-tissue, lung micro-tissue, liver micro-tissue and kidney micro-tissue.
5. The application field is wide. The model can be used in a plurality of fields such as long-term in-vitro tissue culture, drug discovery, drug screening and the like, and has wide application prospects in the aspects of exploring cell differentiation, tissue culture, drug metabolism toxicity and the like.
Drawings
FIG. 1 is a schematic view of an in vitro micro-tissue co-culture well plate according to the present utility model;
FIG. 2 is an exploded view of the culture unit of the present utility model;
FIG. 3 is a top view of the culture unit of the present utility model;
FIG. 4 is a front sectional view of the culture unit of the present utility model;
FIG. 5 is a left side cross-sectional view of the culture unit of the present utility model.
Description of the reference numerals
1-reservoir, 2-culture well, 3-loading well, 4-microfluidic channel, 41-first microfluidic channel, 42-second microfluidic channel, 43-third microfluidic channel, 5-loading microfluidic channel.
Detailed Description
The following description of at least one exemplary embodiment is merely exemplary in nature and is in no way intended to limit the utility model, its application, or uses. All other embodiments, which can be made by those skilled in the art based on the embodiments of the utility model without making any inventive effort, are intended to be within the scope of the utility model.
Referring to fig. 1-5, an in vitro micro-tissue co-culture well plate comprises a well plate body on which at least one set of co-culture units is arranged, each set of culture units comprising:
the liquid storage tank 1 is open at one end and is arranged on the upper surface of the pore plate body;
a culture hole 2, one end of which is open and is arranged on the upper surface of the pore plate body, and the other end of which is communicated with the liquid storage tank 1 through a micro-flow channel 4;
when the experimenter adds the micro-tissue cultured in vitro into the culture well 2 containing the culture medium, the micro-tissue falls into the bottom small hole area due to the action of gravity, thereby limiting the micro-tissue to culture in a fixed range. The design utilizes a physical method to effectively limit the tissue culture area, avoids the camera from scanning a plurality of unnecessary fields of view when high-flux imaging is carried out, and greatly improves the imaging efficiency.
The sample adding hole 3 is open at one end and is arranged on the upper surface of the pore plate body, and the other end is communicated with the culture hole 2 through the microfluidic channel 4;
after the experimenter seeds the micro-tissue into the culture hole 2, the hydrogel and the cell suspension forming the micro-environment can be added into the culture hole 2 through the sample adding hole 3 to form a complex tissue culture micro-environment together with the micro-tissue.
The experimenter can add different types of cells through the loading well 3, such as: mesenchymal cells, endothelial cells, immune cells, etc., thereby mimicking the interaction between tissues and different cells. The application can be used for constructing various research works including tissue culture, vascular microenvironment construction, tumor migration, immunotherapy and the like.
The experimenter can add different types of extracellular matrix through the loading well 3, such as: collagen, matrigel, fibrin, laminin, etc., thereby exploring the effects of different types of extracellular matrix on the growth of micro-tissues.
The liquid storage tanks 1, the culture holes 2 and the sample adding holes 3 are arranged in pairs, each culture hole 2 is provided with the sample adding hole 3, and the liquid storage tanks 1 are arranged on the outer sides of the culture holes 2; each pair of the liquid storage tanks 1 and each pair of the culture wells 2 are communicated through one microfluidic channel 4.
By adding a larger amount of culture medium into the liquid storage groove 1 on one side of the two sides of the culture hole 2, adding a smaller amount of culture medium into the liquid storage groove 1 on the other side, and flowing the culture medium from the side with more culture medium into the side with less culture medium through the micro-flow channel 4 under the action of gravity so as to simulate the flowing condition of blood in a body, thereby forming a dynamic culture environment.
The number of culture wells 2, the height of the microfluidic channel 4, and the shear force of fluid flowing through the culture wells 2 are adjustable. Based on the design of the commercialized culture pore plate structure, the high-throughput culture device is suitable for high-throughput culture, high-throughput sample loading and high-throughput pipetting and pipetting, and the arrangement is suitable for high-throughput printing of various biological 3D printers.
The culture hole 2 and the sample adding hole 3 are funnel-shaped with big top and small bottom, and the inclination angle of the funnel-shaped is 45-75 degrees.
The funnel type inclination angle is 70 degrees.
The microfluidic channel 4 includes a first microfluidic channel 41, a second microfluidic channel 42, and a third microfluidic channel 43, where the first microfluidic channel 41 and the second microfluidic channel 42 are disposed on opposite sides of the third microfluidic channel 43.
The sample loading microfluidic channels 5 are connected to the joints of the first microfluidic channel 41, the third microfluidic channel 43, the second microfluidic channel 42 and the third microfluidic channel 43.
The liquid storage tank 1 is columnar.
The aperture of the sample adding hole 3 is smaller than that of the culture hole 2. The upper part of the well has a pore diameter of 1-4mm, preferably 2.5mm.
The center-to-center distance between the liquid storage tank 1 and the culture hole 2 is 8-10mm.
The working process of the utility model is described in detail below: when the utility model is used, the culture medium is added into the liquid storage tank 1, and the pore plate is inclined, so that the culture medium infiltrates all the culture holes 2. Subsequently, the different micro-tissues are transferred to the culture wells 2, respectively, and the micro-tissues fall into the imaging microwells under the culture wells 2 by gravity. And placing the pore plate on a shaking table, and setting the inclined angle and frequency of the shaking table according to the requirements to construct a dynamic co-culture model.
Co-culture in complex in vitro microenvironment: before the culture medium is not added, the micro-tissue is moved into the culture hole 2 (manual, 3D printing and the like) and then the cell matrix, the cell suspension and the like which are required to be added are injected through the sample adding hole 3, so that the added sample and the micro-tissue can be ensured to be on the same plane, and the subsequent imaging is convenient. Subsequently, the medium is fed through the reservoir 1 and the plate is tilted so that the medium wets all the culture wells 2. And placing the pore plate on a shaking table, and setting the inclination angle and frequency of the shaking table according to the requirements to construct the dynamic complex in-vitro micro-environment co-culture model.
Pipetting: when the liquid is transferred and changed to the cultured sample, firstly, the liquid in the liquid storage tank 1 is sucked away through the liquid transfer gun, and the culture medium remained in the culture hole 2 can be sucked by the liquid transfer gun head close to the funnel-shaped side wall, so that the micro-tissue cultured in the liquid transfer gun head is prevented from being sucked away.
And (3) liquid adding: when the liquid is added, firstly, the culture medium is added into the liquid storage tank 1, then the liquid is added into the culture hole 2, the liquid-transferring gun is abutted against the funnel-shaped inner wall of the culture hole 2, and the culture medium is slowly added, so that the addition of the culture medium in the culture chamber is completed.
According to the utility model, different amounts of culture mediums are added into the liquid storage tanks 1 at two sides of the culture hole 2, and the flow direction of the culture mediums is directionally driven by utilizing the action of gravity, so that the multi-tissue combined dynamic co-culture in-vitro model is constructed. The funnel-shaped structure of the culture hole 2 can limit the micro-tissue to an imaging area at the bottom, so that high-flux and high-connotation imaging is facilitated. Meanwhile, one side of each of the culture wells 2 is provided with a loading well 3, and a user can insert a culture well through the loading well 3 comprising: biological matrices, cell suspensions, etc., to construct complex in vitro culture microenvironments. The well plate is designed based on the well plate structure which is commercially available at present, is simple and convenient to use, and allows high-throughput culture. Therefore, the pore plate can be widely used for in-vitro micro-tissue dynamic co-culture, and has wide application prospects in the fields of tissue culture, drug discovery and the like.
The above description is only of the preferred embodiments of the present utility model and is not intended to limit the present utility model, but various modifications and variations can be made to the present utility model by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present utility model should be included in the protection scope of the present utility model.
Claims (8)
1. An in vitro micro-tissue co-culture well plate, comprising a well plate body on which at least one group of co-culture units is arranged, each group of culture units comprising:
a liquid storage tank (1) with one end open and arranged on the upper surface of the pore plate body;
a culture hole (2) with one end open and arranged on the upper surface of the pore plate body and the other end communicated with the liquid storage tank (1) through a micro-flow channel (4);
a sample adding hole (3) with one end open and arranged on the upper surface of the pore plate body and the other end communicated with the culture hole (2) through a micro-flow channel (4); and
The liquid storage tanks (1), the culture holes (2) and the sample adding holes (3) are all arranged in pairs, each culture hole (2) is provided with the sample adding hole (3), and the liquid storage tanks (1) are arranged on the outer sides of the culture holes (2); each pair of liquid storage tanks (1) and each pair of culture holes (2) are communicated through a micro-flow channel (4).
2. The in vitro micro-tissue co-culture well plate according to claim 1, wherein the culture well (2) and the sample application well (3) are funnel-shaped with large top and small bottom, and the funnel-shaped inclination angle is 45-75 degrees.
3. The in vitro micro-tissue co-culture well plate according to claim 2, wherein the funnel-shaped inclination angle is 70 degrees.
4. An in vitro micro-tissue co-culture well plate according to claim 1, wherein the micro-fluidic channel (4) comprises a first micro-fluidic channel (41), a second micro-fluidic channel (42) and a third micro-fluidic channel (43), the first micro-fluidic channel (41) and the second micro-fluidic channel (42) being arranged opposite to each other on opposite sides of the third micro-fluidic channel (43).
5. The in vitro micro-tissue co-culture well plate according to claim 4, wherein the connection parts of the first micro-flow channel (41) and the third micro-flow channel (43) and the second micro-flow channel (42) and the third micro-flow channel (43) are connected with a sample adding micro-flow channel (5).
6. An in vitro micro-tissue co-culture well plate according to claim 1, wherein the reservoir (1) is cylindrical.
7. An in vitro micro-tissue co-culture well plate according to claim 1, wherein the pore size of the loading well (3) is smaller than the pore size of the culture well (2).
8. An in vitro micro-tissue co-culture well plate according to claim 1, wherein the centre-to-centre spacing of the reservoir (1) and the culture well (2) is 8-10mm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202320667080.0U CN219470073U (en) | 2023-03-30 | 2023-03-30 | In-vitro micro-tissue co-culture pore plate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202320667080.0U CN219470073U (en) | 2023-03-30 | 2023-03-30 | In-vitro micro-tissue co-culture pore plate |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN219470073U true CN219470073U (en) | 2023-08-04 |
Family
ID=87433589
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202320667080.0U Active CN219470073U (en) | 2023-03-30 | 2023-03-30 | In-vitro micro-tissue co-culture pore plate |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN219470073U (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117965272A (en) * | 2024-03-28 | 2024-05-03 | 北京大学人民医院 | Microfluidic chip for bacterial culture, bacterial culture system and bacterial culture method |
-
2023
- 2023-03-30 CN CN202320667080.0U patent/CN219470073U/en active Active
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117965272A (en) * | 2024-03-28 | 2024-05-03 | 北京大学人民医院 | Microfluidic chip for bacterial culture, bacterial culture system and bacterial culture method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Bruzewicz et al. | Fabrication of a modular tissue construct in a microfluidic chip | |
| US9068281B2 (en) | Microfluidic chip for high-throughput perfusion-based three-dimensional cell culture | |
| Tehranirokh et al. | Microfluidic devices for cell cultivation and proliferation | |
| CN219470073U (en) | In-vitro micro-tissue co-culture pore plate | |
| US11680241B2 (en) | Perfusion enabled bioreactors | |
| CN111051494A (en) | 3D cell culture vessel for manual or automatic media exchange | |
| JP7450269B2 (en) | Perfusable bioreactor | |
| CN110551681B (en) | Micro-fluidic chip for simulating embryo implantation angiogenesis and preparation method and application thereof | |
| CN219972330U (en) | Cell culture unit and cell culture device | |
| Ota et al. | Microtechnology-based three-dimensional spheroid formation | |
| CN105695328A (en) | Co-culture device for cell culture and cell-cell interaction and using method of co-culture device | |
| CN108641931A (en) | A kind of digitlization microarray organ chip and its application | |
| CN116240108A (en) | Four-channel organ chip and application and use method thereof | |
| WO2022267247A9 (en) | Culture apparatus | |
| CN113278525B (en) | Stem cell ball or tumor ball culture device and culture method | |
| CN117229917A (en) | Pump-driving-free organoid chip | |
| CN107236668B (en) | Microfluidic chip for breast cancer stem cell culture and drug analysis | |
| CN116478818B (en) | Cell culture unit, device, application and culture method | |
| CN117229918A (en) | Pump-free driving liquid pouring device and method | |
| CN115109703A (en) | Organ chip model | |
| CN117980463A (en) | Cell culture apparatus, cell culture method using the same, and cell culture incubator comprising the same | |
| CN116286342A (en) | Human cerebral cortex-like chip, method for 3D printing human cerebral cortex-like chip and application | |
| CN211999755U (en) | Micro-fluidic chip | |
| CN111647509A (en) | Sitting drop type cell ball culture chip and its use method | |
| JP6937382B2 (en) | Column array and how to create a column array |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant |