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Culture medium for establishing pig iPS cell line and culture method thereof

A culture method and culture medium technology, applied in the field of medium for establishing pig iPS cell lines, can solve problems such as immune rejection, and achieve the effect of neat edges

Inactive Publication Date: 2013-10-02
NORTHWEST A & F UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the acquisition of ES cells, especially human ES cells, has caused many ethical controversies
In addition, the application of ES cells to clinical treatment will also face the problem of immune rejection

Method used

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  • Culture medium for establishing pig iPS cell line and culture method thereof
  • Culture medium for establishing pig iPS cell line and culture method thereof
  • Culture medium for establishing pig iPS cell line and culture method thereof

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Experimental program
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Effect test

Embodiment Construction

[0033] The present invention will be further described in detail below in conjunction with specific embodiments, which are explanations of the present invention rather than limitations.

[0034] A method for obtaining porcine iPS cells. After introducing human OCT4, SOX2, KLF4 and c-MYC into PEF through retroviral vector pMXs, they are cultured on induction medium, which can be efficiently produced under feeder-free conditions. Induce PEF to form porcine iPS cells with an induction efficiency of 0.54%.

[0035] The induction medium is: DMEM+15%FBS+hLIF (10ng / ml)+bFGF (10ng / ml)+CHIR99021 (5μM)+SB431542 (2μM)+hBMP4 (10ng / ml)

[0036] The formed porcine iPS cells are positive for alkaline phosphatase (AP) staining, express Nanog, Sox2, SSEA-1, 4 and other pluripotency markers, and can achieve three germ layer differentiation in vitro and in vivo.

[0037] Specifically, a culture method for establishing a porcine iPS cell line, the operation process is as follows:

[0038] 1) Cu...

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Abstract

The invention discloses a culture medium for establishing a pig iPS cell line and a culture method thereof. A typical pig iPS cell is obtained in the ninth day through transfection by four transcription factors of OCT4, SOX2, KLF4 and c-MYC and induction culture of the culture medium, and the pig iPS cell can be obtained efficiently. The obtained pig iPS cell clone is flat clone, has a regular edge and is similar to the ES cellular morphology of the human body. According to the pig iPS cell line, the pig iPS cell through subculture keeps the undifferentiated state, shows positive in the AP dyeing displaying result and has a pluripotency mark, and the differentiated cell in vitro expresses NCSTN (entoderm), NESTIN (ectoderm) and DESMIN (mesoblast).

Description

technical field [0001] The invention belongs to the technical field of induced pluripotent stem cells, and relates to a culture medium for establishing pig iPS cell lines and a culture method thereof. Background technique [0002] Pluripotent stem cells (pluripotent stem cells) refer to a type of cells that have the potential to form all types of cells in humans and animals and can continuously renew themselves. Due to the multi-directional differentiation potential and self-renewal characteristics of pluripotent stem cells, it has its unique role and advantages in the research of cell replacement therapy, gene therapy, developmental biology research, pharmacology and toxicology. Currently, the most well-known pluripotent stem cells are embryonic stem cells (ES cells), which are generally cell lines established from the inner cell mass of blastocysts under appropriate in vitro culture conditions. However, the acquisition of ES cells, especially the acquisition of human ES c...

Claims

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Application Information

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IPC IPC(8): C12N15/867C12N5/0735
Inventor 张仕强郭燕杰王华岩
Owner NORTHWEST A & F UNIV
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