Cell culture plate coated with Matrigel as well as preparation method and application of cell culture plate

A cell culture, culture plate technology, applied in tissue cell/virus culture devices, biochemical equipment and methods, animal cells, etc., can solve the problems of complex operation, uneven coating, etc. The effect of falling off and easily forming a group

Inactive Publication Date: 2013-05-08
黑龙江省重生生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods all have the disadvantages of complex operation and uneven coating.

Method used

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  • Cell culture plate coated with Matrigel as well as preparation method and application of cell culture plate
  • Cell culture plate coated with Matrigel as well as preparation method and application of cell culture plate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1 is coated with the preparation of the cell culture plate of Matrigel

[0023] (1) Soak polystyrene 6-well cell culture plates in absolute ethanol for 40 min, rinse with deionized water, dry in vacuum, and set aside.

[0024] (2) Measure 50ml of deionized water, add 0.2g of hydroxysuccinimide (NHS) and 0.5g of 2-morpholineethanesulfonic acid hydrate (MES), mix and stir for more than half an hour, filter and sterilize, and store Store at 4°C for later use.

[0025] (3) Weigh 1g of carbodiimide (EDC) and dissolve it in 99ml of deionized water to form an EDC solution with a mass concentration of 1%, filter and sterilize for later use.

[0026] (4) Put the cleaned culture plate into the plasma generator, evacuate to 1Pa, feed oxygen to 50Pa, adjust the discharge power to 60W, and conduct radio frequency discharge for 1min.

[0027] (5) In the culture plate obtained in step (4) under vacuum, add 3ml of the mixed solution prepared in step (2) to each hole, and t...

Embodiment 2

[0029] Embodiment 2 is coated with the preparation of the cell culture plate of Matrigel

[0030] (1) Soak polystyrene 6-well cell culture plates in absolute ethanol for 40 min, rinse with deionized water, dry in vacuum, and set aside.

[0031] (2) Measure 50ml of deionized water, add 0.1g of hydroxysuccinimide (NHS) and 0.5g of 2-morpholineethanesulfonic acid hydrate (MES), mix and stir for more than half an hour, filter and sterilize, and store Store at 4°C for later use.

[0032] (3) Weigh 5g of carbodiimide (EDC) and dissolve it in 95ml of deionized water to form an EDC solution with a mass concentration of 5%, filter and sterilize it for later use.

[0033] (4) Put the cleaned culture plate into the plasma generator, evacuate to 0.5Pa, feed oxygen to 100Pa, adjust the discharge power to 100W, and conduct radio frequency discharge for 1min.

[0034] (5) In the culture plate obtained in step (4) under vacuum, add 3ml of the mixed solution prepared in step (2) to each hole...

Embodiment 3

[0036] Embodiment 3 is coated with the preparation of the cell culture plate of Matrigel

[0037] (1) Soak a polystyrene 24-well cell culture plate in absolute ethanol for 40 minutes, rinse with deionized water, dry in vacuum, and set aside.

[0038] (2) Measure 50ml of deionized water, add 0.05g of hydroxysuccinimide (NHS) and 0.5g of 2-morpholineethanesulfonic acid hydrate (MES), mix and stir for more than half an hour, filter and sterilize, and store Store at 4°C for later use.

[0039] (3) Weigh 3g of carbodiimide (EDC) and dissolve it in 97ml of deionized water to form an EDC solution with a mass concentration of 3%, filter and sterilize for later use.

[0040](4) Put the cleaned culture plate into the plasma generator, evacuate to 1Pa, feed oxygen to 200Pa, adjust the discharge power to 10W, and conduct radio frequency discharge for 5min.

[0041] (5) In the culture plate obtained in step (4) under vacuum, add 0.5ml of the mixed solution prepared in step (2) to each ho...

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Abstract

The invention discloses a cell culture plate coated with Matrigel as well as a preparation method and an application of the cell culture plate. A carboxy surface is formed on the surface of a cell culture plate by adopting a plasma spraying technology, the Matrigel is connected to the surface of the polystyrene cell culture plate or culture dish in a form of a covalent bond by adopting a chemical riveting technology, and the cell culture plate or culture dish coated with the Matrigel is formed. Because a chemical combined surface treatment mode is adopted on the surface of a base material of the culture plate, the Matrigel on the surface is not dissolved. Compared with a physically coated cell culture plate, the cell culture plate has the advantage of long preservation time. The culture plate (dish) is applicable to culture of primary cells and a few cells which are difficult in attaching to walls, easily cluster, easily drop and are slow in growth such as neurons, LNCaP and PC-12; and by adopting the culture plate (dish), the cells attach to the walls firmly and extend fully, and a stable growth state is obtained.

Description

technical field [0001] The invention relates to a cell culture plate coated with Matrigel, a preparation method thereof and an application in cell culture, belonging to the technical field of cell culture. Background technique [0002] Cell culture is a necessary prerequisite for the development of biological science research, cell therapy and tissue engineering. Cell culture plate (dish) is the basic device for cell culture. Currently, the cell culture plate is mainly made of polystyrene or polyethylene. It is suitable for Adhesion and growth of a variety of cells. With the development of cell biology, the performance requirements of cell culture plates are getting higher and higher. Ordinary cell culture plates are difficult for some primary cells to adhere to the wall, easy to agglomerate, easy to fall off, and slow-growing cells to proliferate. not effectively. In order to improve the adhesion and proliferation of these cells, in recent years, people have coated polyly...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/04C12N5/00C12N5/079C12N5/0793C12N5/09
Inventor 李萌
Owner 黑龙江省重生生物科技有限公司
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