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Serum-free and feeder-layer-free culture medium for effectively inhibiting stem cell differentiation and culture method

A serum-free medium and stem cell culture technology, applied in cell culture active agents, non-embryonic pluripotent stem cells, biochemical equipment and methods, etc., can solve problems such as poor application prospects and complex components

Pending Publication Date: 2021-09-14
BIOISLAND LAB +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although activinA is used in this patent to maintain the pluripotency of embryonic stem cells, the patent uses a serum substitute, the composition is complex and the application prospect is not good

Method used

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  • Serum-free and feeder-layer-free culture medium for effectively inhibiting stem cell differentiation and culture method
  • Serum-free and feeder-layer-free culture medium for effectively inhibiting stem cell differentiation and culture method
  • Serum-free and feeder-layer-free culture medium for effectively inhibiting stem cell differentiation and culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] A serum-free feeder-free culture medium for embryonic stem cells or pluripotent stem cells in this embodiment, the formula of which is shown in the table below:

[0044]

[0045]

[0046] Its preparation method is as follows: dissolve each additive component in the medium in the above table according to their respective solubility characteristics, and then add each component to the DMEM / F12 basal medium one by one at 20°C under sterile conditions after filter sterilization. , mix well by pipetting, adjust the osmotic pressure to 340mOsm / kg with sodium chloride, and store the prepared medium at 4°C.

Embodiment 2

[0048] This embodiment provides a serum-free feeder-free culture medium for embryonic stem cells or pluripotent stem cells, the formula of which is shown in the following table:

[0049]

[0050]

[0051] Its preparation method is as follows: dissolve each additive component in the medium in the above table according to their respective solubility characteristics, and then add each component to the DMEM / F12 basal medium one by one at 20°C under sterile conditions after filter sterilization. , mix well by pipetting, adjust the osmotic pressure to 340mOsm / kg with sodium chloride, and store the prepared medium at 4°C.

Embodiment 3

[0053] This embodiment provides a serum-free feeder-free culture medium for embryonic stem cells or pluripotent stem cells, the formula of which is shown in the following table:

[0054]

[0055] Its preparation method is as follows: dissolve each additive component in the medium in the above table according to their respective solubility characteristics, and then add each component to the DMEM / F12 basal medium one by one at 20°C under sterile conditions after filter sterilization. , mix well by pipetting, adjust the osmotic pressure to 340mOsm / kg with sodium chloride, and store the prepared medium at 4°C.

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Abstract

The invention relates to a serum-free and feeder-layer-free culture medium for effectively inhibiting stem cell differentiation and a culture method. The invention also relates to stem cells maintained in an undifferentiated state. The previous culture of the human embryonic stem cells requires feeder layer cells and serum-containing media to maintain the stem cells in an undifferentiated state. The invention finds that if TGF-beta and Activin are added into a culture medium for culturing the stem cells, feeder cells and serum are not needed, and the stem cells can remain in an undifferentiated state indefinitely in at least 10 passages.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a serum-free and feeder-free medium and a culture method for effectively inhibiting stem cell differentiation. Background technique [0002] Cells in many tissues such as skin, blood, and intestinal epithelium in adult animals are short-lived and need to be constantly replaced by corresponding new cells. One of the ways for mature individuals to produce new differentiated cells is to form new differentiated cells through simple multiplication of existing differentiated cells, that is, differentiated cells divide to form two daughter cells of the same type, such as new endothelial cells in blood vessels through generated in this way. However, in the process of differentiation, cells often lose the ability to divide again due to high differentiation, and eventually go to senescence and die. In order to make up for this deficiency, the body also retains a part of undifferenti...

Claims

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Application Information

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IPC IPC(8): C12N5/0735C12N5/074
CPCC12N5/0606C12N5/0696C12N2500/90C12N2501/15C12N2501/998
Inventor 陈东煌陈海佳张兆清姜交华李学家
Owner BIOISLAND LAB
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