CN114907979A - Gridding embedded cell co-culture device and method - Google Patents
Gridding embedded cell co-culture device and method Download PDFInfo
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Abstract
The invention provides a gridding embedded cell co-culture device and a gridding embedded cell co-culture method, wherein the device comprises a dish body, a dish cover, a first culture plate and a second culture plate; the first culture plate and the second culture plate respectively comprise a plurality of culture tanks which are distributed at intervals in a net shape and a plurality of hollow parts which are arranged between the adjacent culture tanks; the hollow part of the first culture plate corresponds to the culture groove position of the second culture plate, and the hollow part of the second culture plate corresponds to the culture groove position of the first culture plate; the height of the side wall of the culture tank is less than that of the side wall of the dish body; the dish cover is matched with the dish body; when the first culture plate and the second culture plate are spliced and matched in a staggered way to form a whole, the culture grooves of the first culture plate and the second culture plate are distributed in a staggered way; the first culture plate and the second culture plate which are spliced in a staggered mode can be accommodated in the dish body. The gridding embedded cell co-culture device and method can realize the free-ratio gridding non-contact co-culture of multiple cells, and have simple production process and high efficiency.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a gridding embedded cell co-culture device and a gridding embedded cell co-culture method.
Background
Tumors are one of the most important factors threatening human life, and thus, intensive research into tumors has been attempted to achieve the elimination of tumors. The tumor is in a highly information microenvironment in the human body, but most of the research on the tumor now researches biological information when tumor cells exist alone, and the research results of the research on the tumor departing from the microenvironment can be different from the conditions of the tumor in the human body. Many drugs have good killing effect in vitro experiments, but the tumor killing effect in vivo is not satisfactory, and the conclusion is further proved. Therefore, in order to establish a tumor culture system which is more in line with the in vivo environment, people can make the in vitro tumor research environment matched with the in vivo tumor environment as much as possible, so that the tumor cells can communicate with each other, and the cell co-culture technology can be developed.
There are two main methods for cell co-culture: direct contact co-culture and non-contact co-culture. The contact co-culture method co-cultures different cells according to different proportions, and the application range in scientific research is not wide because the different cells are not directly contacted and thoroughly separated to carry out the next research and analysis. Another co-culture mode is non-contact co-culture, which separates different cells by different methods to achieve the purpose of being in the same culture system without direct contact. At present, the Chinese patent application numbers are: CN 112920992 a, entitled "method for co-culturing mesenchymal stem cells and nucleus pulposus cells and application", but this co-culture device obviously has the following disadvantages: 1. the price is expensive: the Transwell culture chamber is high in price; 2. co-culture of more than two adherent cells cannot be realized; 3. it is inconvenient to observe the state of the cells in the lower chamber. In addition, the Chinese patent application numbers are: CN 104164365B, entitled "in vitro cell contact type co-culture device and culture operation method thereof", but the co-culture device also has obvious defects: 1. co-culture of more than two adherent cells cannot be achieved. 2. After the co-culture, one cell alone cannot be subjected to subsequent experimental manipulations. In addition, the Chinese patent application numbers are: CN 201545843U, entitled "a non-direct contact cell co-culture device", however, this device could not realize co-culture of more than 2 cells and co-culture at a freely set ratio.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a gridding embedded cell co-culture device and a gridding embedded cell co-culture method, which can realize the free-ratio gridding non-contact co-culture of multiple cells, and have simple production process and high efficiency.
In order to achieve the above object, the present invention provides a gridding embedded cell co-culture device, comprising a dish body, a dish cover, a first culture plate and a second culture plate; the first culture plate and the second culture plate respectively comprise a plurality of culture grooves which are distributed at intervals in a net shape and a plurality of hollow parts which are arranged between the adjacent culture grooves; the hollow part of the first culture plate corresponds to the culture groove position of the second culture plate, and the hollow part of the second culture plate corresponds to the culture groove position of the first culture plate; the height of the side wall of the culture tank is smaller than that of the side wall of the dish body; the dish cover is matched with the dish body; when the first culture plate and the second culture plate are in staggered splicing fit to form a whole, the culture grooves of the first culture plate and the second culture plate are distributed in a staggered manner; the dish body can accommodate the first culture plate and the second culture plate which are spliced in a staggered mode.
Preferably, the first culture plate and the second culture plate are in staggered splicing fit and are rectangular.
Preferably, the culture tank is rectangular.
Preferably, the side wall of the culture tank is 5mm high; the height of the side wall of the dish body is 23 mm.
Preferably, the distance between two adjacent culture grooves of the first culture plate and the second culture plate which are distributed in a staggered way is more than or equal to 2 mm.
Preferably, the dish body, the dish cover, the first culture plate and the second culture plate are formed by 3D printing.
The invention relates to a gridding embedded cell co-culture method based on a gridding embedded cell co-culture device, which comprises the following steps:
s1: a cell paving step;
the step of S1 further includes the steps of:
s11: inserting the first culture plate and the second culture plate together in a staggered manner, and putting the culture plates into the dish body;
s12: adding 200ul of cell suspension to the culture tank, wherein the cell density of the cell suspension is less than 80%; avoid shaking the utensil body hard;
s13: the cell suspension was incubated at 37 ℃ with 5% CO by volume 2 Until the cells in the cell suspension adhere to the wall;
s2: forming a co-culture environment;
the step of S2 further includes the steps of:
s21: discarding the original culture medium in the culture tank;
s22: adding a sufficient amount of fresh culture medium to cover each culture well and to integrate the culture wells;
s23: the cells were incubated at 37 ℃ and 5% CO by volume 2 Incubating for a period of time;
s3: finishing the co-culture environment;
the step of S3 further includes the steps of:
s31: discarding the dish body and the new culture medium in the culture tank;
s32: separating the first culture plate and the second culture plate;
s33: collecting the cells in different culture tanks respectively.
Preferably, in the step S12, the cell suspensions of different cells are set to be cultured at different culture ratios.
Due to the adoption of the technical scheme, the invention has the following beneficial effects:
1. the gridding staggered culture tank structure of the first culture plate and the second culture plate can realize the free-ratio gridding non-contact co-culture of multiple cells, and the production process is simple and has high efficiency.
2. Different plating schemes can be adopted, for example, the surrounding culture is more suitable for the living environment of the cells in the body, the potential defect that the research on the cells is separated from the biological information communication between the cells is avoided, and the repeatability and the stability of the experiment are greatly improved
3. The first culture plate and the second culture plate are provided with a plurality of culture grooves, and culture modes with different proportions can be freely set, so that the culture plate is an ideal cell co-culture model.
4. The interval of 2mm between each culture tank avoids the error caused by impure cells in the small holes due to the mixing of different types of cells caused by cell migration.
5. The most outstanding advantage is that in the practical work, due to the limitation of supporting equipment and technology such as transwell co-culture and the like, the requirement of multi-cell free ratio co-culture cannot be completely met, and the co-culture mode provided by the invention can simulate the real living state of cells in vivo to the maximum extent; meanwhile, the invention is suitable for various adherent cells, and greatly improves the application range, the identification degree and the accuracy of the invention in scientific research.
The multi-cell free-ratio non-contact co-culture technology provided by the invention can be used for in vivo and in vitro research, molecular mechanism research, pharmacological mechanism and pharmacotherapy evaluation research and the like of various cells, and provides a solid tool for researching the field of medical biology research; has wide application prospect and potential social and economic values in basic research and preliminary clinical research of medicaments.
Drawings
FIG. 1 is a schematic structural diagram of a gridding embedded cell co-culture device according to an embodiment of the present invention;
FIG. 2 is a schematic view of a first culture plate according to an embodiment of the present invention;
FIG. 3 is a schematic diagram of a second growth plate according to an embodiment of the present invention.
Detailed Description
The following description of the preferred embodiments of the present invention will be provided in conjunction with the accompanying drawings 1-3, and will make the functions and features of the invention better understood.
Referring to fig. 1 to 3, a gridding embedded cell co-culture device according to an embodiment of the present invention includes a dish body 1, a dish cover, a first culture plate 2 and a second culture plate 3; the first culture plate 2 and the second culture plate 3 respectively comprise a plurality of culture grooves 4 which are distributed at intervals in a net shape and a plurality of hollow parts 5 which are arranged between the adjacent culture grooves 4; the hollow part 5 of the first culture plate 2 corresponds to the culture groove 4 of the second culture plate 3 in position, and the hollow part 5 of the second culture plate 3 corresponds to the culture groove 4 of the first culture plate 2 in position; the height of the side wall of the culture tank 4 is less than that of the side wall of the dish body 1; the dish cover is matched with the dish body 1; when the first culture plate 2 and the second culture plate 3 are inserted and matched in a staggered way to form a whole, the culture grooves 4 of the first culture plate 2 and the second culture plate 3 are distributed in a staggered way; the dish body 1 can accommodate the first culture plate 2 and the second culture plate 3 which are spliced in a staggered mode.
The first culture plate 2 and the second culture plate 3 are in staggered splicing fit to form a rectangle.
The culture tank 4 has a rectangular shape.
The height of the side wall of the culture tank 4 is 5 mm; the height of the side wall of the dish body 1 is 23 mm.
The distance between two adjacent culture grooves 4 which are distributed in a staggered way of the first culture plate 2 and the second culture plate 3 is more than or equal to 2 mm.
The gridding embedded cell co-culture method based on the gridding embedded cell co-culture device of the embodiment of the invention comprises the following steps:
s1: a cell paving step;
the step of S1 further includes the steps of:
s11: inserting the first culture plate 2 and the second culture plate 3 into the dish body 1 in a staggered manner;
s12: 200ul of cell suspension is added into the culture tank 4, and the cell density of the cell suspension is less than 80%; avoid shaking the vessel body 1 with force;
s13: the cell suspension was incubated at 37 ℃ with 5% CO by volume 2 Until the cells in the cell suspension adhere to the wall;
s2: forming a co-culture environment;
the step of S2 further includes the steps of:
s21: discarding the original culture medium in the culture tank 4;
s22: adding enough new culture medium to cover each culture tank 4, and connecting the culture tanks 4 into a whole;
s23: cells were incubated at 37 ℃ with 5% CO by volume 2 Incubating for a period of time;
s3: finishing the co-culture environment;
the step of S3 further includes the steps of:
s31: discarding the new culture medium in the dish body 1 and the culture tank 4;
s32: separating the first culture plate 2 and the second culture plate 3;
s33: the cells in the different culture tanks 4 are collected.
In step S12, cell suspensions of different cells can be set to be cultured at different culture ratios.
Different culture modes, such as surrounding co-culture and cross co-culture, are realized by regulating the arrangement mode of each culture tank 4 of the cells.
Through different culture modes, influence factors of behaviors such as proliferation, differentiation, migration, apoptosis and the like of target cells in a co-culture system can be explored.
After the co-culture is finished, the first culture plate 2 and the second culture plate 3 can be easily separated, and the first culture plate 2 and the second culture plate 3 can be collected respectively for subsequent experiments, such as cell cycle detection, molecular protein detection and the like.
The method provided by the invention not only can realize co-culture of two or more kinds of cells in different proportions, but also can easily avoid errors caused by the separation complexity of a subsequent flow cytometer in contact type co-culture.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (8)
1. A gridding embedded cell co-culture device comprises a dish body, a dish cover, a first culture plate and a second culture plate; the method is characterized in that: the first culture plate and the second culture plate respectively comprise a plurality of culture grooves which are distributed at intervals in a net shape and a plurality of hollow parts which are arranged between the adjacent culture grooves; the hollow part of the first culture plate corresponds to the culture groove position of the second culture plate, and the hollow part of the second culture plate corresponds to the culture groove position of the first culture plate; the height of the side wall of the culture tank is smaller than that of the side wall of the dish body; the dish cover is matched with the dish body; when the first culture plate and the second culture plate are in staggered splicing fit to form a whole, the culture grooves of the first culture plate and the second culture plate are distributed in a staggered manner; the dish body can accommodate the first culture plate and the second culture plate which are spliced in a staggered mode.
2. The meshed embedded cell co-culture device according to claim 1, wherein the first culture plate and the second culture plate are in a rectangular shape by staggered insertion fit.
3. The meshed embedded cell co-culture device according to claim 2, wherein the culture tank has a rectangular shape.
4. The meshed embedded cell co-culture device according to claim 3, wherein the side wall of the culture tank is 5mm high; the height of the side wall of the dish body is 23 mm.
5. The meshed embedded cell co-culture device according to claim 4, wherein the spacing between two adjacent culture grooves of the first culture plate and the second culture plate in the staggered distribution is greater than or equal to 2 mm.
6. The meshed embedded cell co-culture device according to claim 5, wherein the dish body, the dish lid, the first culture plate and the second culture plate are molded by 3D printing.
7. A gridding embedded cell co-culture method based on the gridding embedded cell co-culture device according to claim 1, comprising the steps of:
s1: a cell paving step;
the step of S1 further includes the steps of:
s11: inserting the first culture plate and the second culture plate together in a staggered manner, and putting the culture plates into the dish body;
s12: adding 200ul of cell suspension to the culture tank, wherein the cell density of the cell suspension is less than 80%; avoid shaking the utensil body hard;
s13: the cell suspension was incubated at 37 ℃ with 5% CO by volume 2 Until the cells in the cell suspension adhere to the wall;
s2: forming a co-culture environment;
the step of S2 further includes the steps of:
s21: discarding the original culture medium in the culture tank;
s22: adding a sufficient amount of fresh culture medium to cover each culture well and to integrate the culture wells;
s23: the cells were incubated at 37 ℃ and 5% CO by volume 2 Incubating for a period of time;
s3: finishing the co-culture environment;
the step of S3 further includes the steps of:
s31: discarding the dish body and the new culture medium in the culture tank;
s32: separating the first culture plate and the second culture plate;
s33: collecting the cells in different culture tanks respectively.
8. The method for co-culturing embedded cells in a grid structure according to claim 7, wherein in the step S12, the cell suspensions of different cells are cultured at different culture ratios.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112920992A (en) * | 2021-01-12 | 2021-06-08 | 常州市第二人民医院 | Method for co-culturing mesenchymal stem cells and nucleus pulposus cells and application |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112920992A (en) * | 2021-01-12 | 2021-06-08 | 常州市第二人民医院 | Method for co-culturing mesenchymal stem cells and nucleus pulposus cells and application |
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