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Mouse embryonic stem cell (ESC) induction culture solution for improving 2C-like cells and mouse ESC induction culture method

A 2c-like, mouse embryo technology, applied in the direction of cell culture active agent, embryonic cells, culture process, etc.

Active Publication Date: 2020-02-18
INNER MONGOLIA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the maintenance and self-renewal of 2C-like cells in mouse ESCs has not been maintained under chemically defined culture conditions so far.

Method used

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  • Mouse embryonic stem cell (ESC) induction culture solution for improving 2C-like cells and mouse ESC induction culture method
  • Mouse embryonic stem cell (ESC) induction culture solution for improving 2C-like cells and mouse ESC induction culture method
  • Mouse embryonic stem cell (ESC) induction culture solution for improving 2C-like cells and mouse ESC induction culture method

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Embodiment Construction

[0043] Below in conjunction with accompanying drawing and specific embodiment the present invention is described in further detail:

[0044] The mouse embryonic stem cell induction medium for improving 2C-like cells of the present invention, every 500ml induction medium N2B27+2i / LIF+RA consists of:

[0045] N2 cell culture supplement (purchased from Gibco, product number: 17502-048) 2-3 ml

[0046] B27 cell culture supplement (purchased from Gibco, item number: 17504-044) 4-6 ml

[0047] Bovine serum albumin (purchased from Sigma, product number: A3311) 20-30 mg

[0048] Non-essential amino acids (purchased from Gibco, catalog number: 11140-035) 4-6 ml

[0049] L-glutamine (purchased from Gibco, catalog number: 35050-061) 4-6 ml

[0050] β-mercaptoethanol (purchased from Gibco, catalog number: 21985-023) 0.5-2 ml

[0051] Penicillin streptomycin (purchased from Sigma, product number: 010M0650) 4-5 ml

[0052] PD0325901 (purchased from Miltenyi Biotec, catalog number: 130-...

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Abstract

The invention discloses a mouse embryonic stem cell (ESC) induction culture solution for improving 2C-like cells and a mouse ESC induction culture method. The method comprises the steps that: first, fetal bovine serum is removed from a mouse ESCs culture system, so that mouse ESCs still maintain original pluripotency characteristics; and second, retinoic acid (RA) is utilized for increasing the proportion of the 2C-like cells in the mouse ESCs in a culture solution without containing the fetal bovine serum, and the result is increased from 0.2% of a control group to 3.3%. The 2C-like cell proportion in the mouse ESCs is increased by using a specific induction method, the 2C-like gene expression level and protein level of the mouse ESCs in the culture solution without containing the fetal bovine serum are verified by experimental means, and a new thought is provided for research of mammal totipotent stem cells.

Description

technical field [0001] The invention relates to a mouse embryonic stem cell induction culture medium, in particular to a mouse embryonic stem cell induction culture medium for improving 2C-like cells and a mouse embryonic stem cell induction culture method. Background technique [0002] Mammalian zygotic genome activation (ZGA) is a key step in embryogenesis, which occurs at the 1-2 cell stage in mice and the 4-8 cell stage in humans, and this stage is important for early embryonic development and the formation of totipotency in mammals significance. [0003] Mouse ESCs contain a fraction (less than 1%) of heterogeneous cells that resemble 2-cell stage embryonic-like cells (2C-like cells). 2C-like cells can not only express ZGA transcripts, but also have epigenetic features similar to 2-cell embryos. In addition, 2C-like cells in mouse ESCs can contribute to both embryonic and extraembryonic tissues during in vivo development. Therefore, this part of 2C-like cells is a pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0735C12N5/073C12N5/10
CPCC12N5/0606C12N5/0603C12N2501/235C12N2501/998C12N2500/32C12N2500/44C12N2501/385C12N2501/727C12N2510/00
Inventor 包斯琴吴宝江王艳秋李喜和
Owner INNER MONGOLIA UNIVERSITY
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