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Serum-free medium of embryonic stem cells, and its application

A serum-free medium, embryonic stem cell technology, applied in the field of embryonic stem cell culture, to achieve the effect of maintaining self-renewal and clear components

Inactive Publication Date: 2014-02-05
苏州依科赛生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the serum-free medium of embryonic stem cells containing CHIR99021 and XAV939.

Method used

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  • Serum-free medium of embryonic stem cells, and its application
  • Serum-free medium of embryonic stem cells, and its application
  • Serum-free medium of embryonic stem cells, and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Embryonic stem cell serum-free medium of the present invention (1)

[0029] The medium is composed of the following components: CHIR99021, XAV939, B27 / N2 and DMEM / F12. The specific material composition and working concentration of each component are:

[0030] (1) CHIR99021 0.85 μmol / L,

[0031] (2) XAV939 1.02 μmol / L,

[0032] (3) Additive N2:

[0033]

[0034] (4) Additive B27:

[0035]

[0036] (5) DMEM / F12:

[0037]

[0038]

[0039]

[0040] According to the above components, the culture medium was prepared by conventional methods.

Embodiment 2

[0041] Example 2 Embryonic stem cell serum-free medium of the present invention (2)

[0042] The medium is composed of the following components: CHIR99021, XAV939, B27 / N2 and DMEM / F12. The specific material composition and working concentration of each component are:

[0043] (1) CHIR99021 0.80μmol / L,

[0044] (2) XAV939 1.08 μmol / L,

[0045] (3) Additive N2:

[0046]

[0047] (4) Additive B27:

[0048]

[0049] (5) DMEM / F12:

[0050]

[0051]

[0052]

[0053] According to the above components, the culture medium was prepared by conventional methods.

Embodiment 3

[0054] Example 3 Embryonic stem cell serum-free medium of the present invention (3)

[0055] The medium is composed of the following components: CHIR99021, XAV939, B27 / N2 and DMEM / F12. The specific material composition and working concentration of each component are:

[0056] (1) CHIR99021 0.95 μmol / L,

[0057] (2) XAV939 0.95 μmol / L,

[0058] (3) Additive N2:

[0059]

[0060] (4) Additive B27:

[0061]

[0062] (5) DMEM / F12:

[0063]

[0064]

[0065]

[0066] According to the above components, the culture medium was prepared by conventional methods.

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Abstract

The invention relates to a serum-free medium of embryonic stem cells, and its application. The medium comprises a micro-molecular inhibitor CHIR99021, a micro-molecular inhibitor XAV939, an additive B27 / N2 and a basic medium DMEM / F12. The serum-free medium of embryonic stem cells, which contains no animal source substances and has a determined composition, is obtained by using the micro-molecular inhibitor CHIR99021, the micro-molecular inhibitor XAV939 and the B27 / N2 additive having a clear composition to substitute serum and feeder cells in a traditional embryo hepatocyte medium. Experiments prove that the medium can maintain the self updating of human embryonic stem cells, and maintains the undifferentiated state and totipotency of the embryonic stem cells; and the medium has a clear composition, and proteins and other components contained in the medium are from human recombinant proteins or are obtained through chemical synthesis, so there is no embryonic stem cell pollution, thereby no corresponding immune response is excited, and no immune immunological rejection is initiated.

Description

technical field [0001] The invention relates to the cultivation of embryonic stem cells, in particular to a serum-free culture medium for embryonic stem cells and its application. Background technique [0002] Embryonic Stem Cells (ES Cells) are a kind of totipotent stem cells selected from early embryonic inner cell mass (Inner cell mass, ICM) or primordial germ cells (Primordial germ cells, PGCs) through in vitro differentiation inhibition culture. The morphological characteristics of embryonic stem cells are very similar to those of early embryonic cells. They have stable self-renewal ability and can maintain the normal diploid chromosome structure after long-term subculture in vitro. At the same time, embryonic stem cells also have very strong differentiation ability. Under conditional culture, it can proliferate and differentiate into more than 200 cell types throughout the body. Therefore, embryonic stem cells can provide an ideal biological platform for scientific re...

Claims

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Application Information

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IPC IPC(8): C12N5/0735
Inventor 吕万革郭亚楠陈旭陈刚
Owner 苏州依科赛生物科技股份有限公司
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