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Separation culture method and culture medium for chicken embryonic stem cells

A chicken embryonic stem cell and culture medium technology, applied in cell dissociation methods, embryonic cells, cell culture active agents, etc., can solve the problems of slow cell proliferation, easy differentiation, and low cloning rate.

Inactive Publication Date: 2019-09-27
JILIN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method still has the disadvantages of low cloning rate, slow cell proliferation, and easy differentiation, so it is necessary to do further research on the isolation and culture method of chicken embryonic stem cells

Method used

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  • Separation culture method and culture medium for chicken embryonic stem cells
  • Separation culture method and culture medium for chicken embryonic stem cells
  • Separation culture method and culture medium for chicken embryonic stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1. Experimental animals

[0029] Sea blue brown layer fresh eggs and 9-10d hatched chicken embryos: Collected from Mailong Animal Husbandry and Xinhui Breeding Cooperative in Gongzhuling City, Jilin Province.

[0030] 2. Preparation of chicken embryo fibroblast culture medium

[0031] High glucose DMEM containing the following solutes: 10% fetal bovine serum (volume ratio), 1% penicillin / streptomycin (volume ratio).

[0032] 3. Preparation of basal medium

[0033] High-sugar DMEM containing the following solutes: 10% FBS (volume ratio), 2% chicken serum (volume ratio), 1mmol / L sodium pyruvate, 2mmol / L L-glutamine, 1% non-essential amino acids (volume ratio) , 5.5×10 -5 mmol / Lβ-mercaptoethanol, 1% penicillin / streptomycin (volume ratio).

[0034] 4. Preparation of chicken embryonic stem cell complete medium

[0035] Add to the above basal medium: 10 ng / mL bFGF, 0.1 ng / mL mLIF, 5 ng / mL hSCF, 10 ng / mL hIGF, 20 ng / mL conalbumin, 1 μmol / L nucleotide adenosine, 1 ng / mL hI...

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Abstract

The invention discloses a separation culture method and a culture medium for chicken embryonic stem cells. The culture medium is obtained by adding a basic fibroblast growth factor, a stem cell factor, a leukaemia inhibitory factor, an insulin-like growth factor-1, conalbumin, adenosine nucleotide, recombinant interleukin-6 and recombinant interleukin-11 into a basic culture medium. According to the separation culture method and the culture medium for the chicken embryonic stem cells, the chicken embryonic stem cells can be cultured stably for a long term in vitro and have good augmentation and renewal capabilities by adding the cell factors, such as the insulin-like growth factor-1, the conalbumin, the adenosine nucleotide, IL-6 and IL-11, exogenously for the first time.

Description

technical field [0001] The invention relates to the technical field of stem cells, in particular to a method for separating and culturing chicken embryonic stem cells and a culture medium. Background technique [0002] Embryonic stem cells are a type of cells isolated from the inner cell mass of early embryos. They not only have the same unlimited proliferation, self-renewal and multi-directional differentiation potential as other stem cells, but also have developmental totipotency. Under certain conditions, they can be induced Differentiate into almost all cell types of the inner, middle and outer three germ layers of the body. In 1981, based on the study of epithelial cells, Evans and Kaufmanz first cultured delayed implantation mouse early embryos in vitro, isolated mouse embryonic stem cells for the first time, and established embryonic stem cell lines. Embryonic stem cells have the ability of germline transmission and provide good seed resources for gene modification, ...

Claims

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Application Information

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IPC IPC(8): C12N5/0735
CPCC12N5/0606C12N2500/40C12N2501/105C12N2501/115C12N2501/125C12N2501/2306C12N2501/2311C12N2501/998C12N2509/00
Inventor 李欣赵中利于永生金海国曹阳张立春朴庆林吕礼良
Owner JILIN ACAD OF AGRI SCI