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Co-culture device for cell culture and cell-cell interaction and using method of co-culture device

A cell culture and co-culture technology, which is applied in the direction of tissue cell/virus culture devices, sterilization methods, biochemical equipment and methods, etc., can solve problems such as the inability to rule out gravity interference, limit the influence of tumor cell functions, and avoid The effect of cumbersome separation steps, wide applicability and low cost

Inactive Publication Date: 2016-06-22
张姝 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The use of traditional Transwell chambers to realize the co-culture of tumor cells into spheres and other cells has the following problems: (1) Due to the special requirements of tumor three-dimensional sphere culture, tumor cells can only be confined to the lower layer of Transwell chambers, thus limiting the ability of other cells to grow into spheres. Research on some functional effects of spherical tumor cells; (2) Transwell chamber is a structure for cell culture in the upper and lower layers, and the defect of gravity interference cannot be ruled out

Method used

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  • Co-culture device for cell culture and cell-cell interaction and using method of co-culture device
  • Co-culture device for cell culture and cell-cell interaction and using method of co-culture device
  • Co-culture device for cell culture and cell-cell interaction and using method of co-culture device

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Experimental program
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Effect test

Embodiment 1

[0035] S1. Carrying out autoclaving and ultraviolet irradiation sterilization on the co-cultivation device;

[0036] S2. Insert a detachable semi-permeable membrane spacer 6 with an aperture size of 3 μm in the middle slot, and insert detachable spacers 7a and 7b into the slots on both sides;

[0037] S3. Adding the tumor cell suspension into the cell culture chamber 3 coated with matrigel under aseptic operation, allowing the tumor cells to sink in the matrigel, and stably growing and culturing in the cell culture chamber 3 for 3 days;

[0038] S4. On the 4th day, another cell culture chamber 4 was coated with poly-lysine, and another adherent cell was planted in the cell culture chamber 4 coated with poly-lysine. Cultured overnight at 37°C to make Adherent growth in cell culture chamber 4;

[0039] S5, detach the detachable spacers 7a and 7b, and co-culture the tumor cells and the adherent cells;

[0040] S6. Continue the co-cultivation, observe the formation of tumor cells ...

Embodiment 2

[0042] S1. Carrying out autoclaving and ultraviolet irradiation sterilization on the co-cultivation device;

[0043] S2. Insert a detachable semi-permeable membrane spacer 6 with an aperture size of 8 μm in the middle slot, and insert detachable spacers 7a and 7b into the slots on both sides;

[0044] S3. Adding the tumor cell suspension into the cell culture chamber 3 coated with matrigel under aseptic operation, allowing the tumor cells to sink in the matrigel, and stably growing and culturing in the cell culture chamber 3 for 3 days;

[0045] S4. On the 4th day, another cell culture chamber 4 was coated with poly-lysine, and another adherent cell was planted in the cell culture chamber 4 coated with poly-lysine. Cultured overnight at 37°C to make Adherent growth in cell culture chamber 4;

[0046] S5, detach the detachable spacers 7a and 7b, and co-culture the tumor cells and the adherent cells;

[0047] S6. Continue the co-cultivation, observe the tumor cell sphere forma...

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Abstract

The invention discloses a co-culture device for cell culture and cell-cell interaction and a using method of the co-culture device. The device comprises a culture plate, wherein the culture plate is provided with at least one culture hole, at least one detachable semipermeable membrane spacing block and detachable spacing blocks; each detachable semipermeable membrane spacing block divides the corresponding culture hole into a plurality of cell culture rooms; and the detachable spacing blocks are positioned on two sides of each detachable semipermeable membrane spacing block. The using method of the device comprises the following steps: inserting the detachable semipermeable membrane spacing blocks and the detachable spacing blocks in the culture holes after sterilization; culturing tumor cells in a segmented cell culture room; culturing adherent cells in another cell culture room; detaching the detachable spacing blocks, and enabling the tumor cells and the adherent cells to be in co-culture; and continuing co-culture, observing a tumor cell balling condition and a migration condition of the balled tumor cells from one cell culture room to the other cell culture room. The co-culture device for cell culture and cell-cell interaction is simple in structure, simple and practical in operation, and visual and clear in effect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a co-cultivation device for cell culture and interaction between cells and its application method. Background technique [0002] The use of ordinary two-dimensional cell culture to study the mechanism of tumor development is very different from the real in vivo environment, such as cell-cell, cell-matrix interaction, cell differentiation, etc. Due to the species differences between animal models and the human body, the characteristics of the human body cannot be well replicated, such as human tumor growth and metastasis, drug treatment response, immune response, and tumor stem cell (TSCs) differentiation. [0003] The three-dimensional cell culture technology is simple and easy to operate, and can simulate the characteristics of the in vivo environment. In 3D culture, single cell types of tumor cells or TSCs can be included, and multiple cell lines such as tumor cells, stroma and im...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/02C12N5/09
CPCC12M23/12C12M25/02C12M37/04C12N5/0693C12N2502/30
Inventor 张姝孙新臣朱晓敏卢倩王远东何国凤严菁菁
Owner 张姝
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