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Novel animal source-free and feed layer-free human pluripotent stem cell culture system

A technology of human pluripotent stem cells and pluripotent stem cells, which is applied in the field of cell culture, can solve problems such as obstacles to clinical application, and achieve the effects of low cost, wide range of sources, and low production cost

Inactive Publication Date: 2012-07-18
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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Problems solved by technology

However, the culture is directly exposed to substances of mouse and bovine origin, which will have great obstacles to the clinical application in the future.

Method used

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  • Novel animal source-free and feed layer-free human pluripotent stem cell culture system
  • Novel animal source-free and feed layer-free human pluripotent stem cell culture system
  • Novel animal source-free and feed layer-free human pluripotent stem cell culture system

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Embodiment Construction

[0043] The novel animal source-free, feeder-free stem cell culture system provided by the present invention mainly includes two main aspects: 1) the extracellular matrix (FCF matrix) on which stem cells grow; 2) the cells that maintain the self-renewal of embryonic stem cells Medium (XF medium).

[0044] 1. Extraction of humanized matrix

[0045] By optimizing the extraction method of Matrigel, we have developed a simple and easy method for extracting extracellular matrix from placenta.

[0046] First, we obtained the placenta of healthy pregnant women who tested negative for hepatitis B, AIDS, and syphilis from the hospital. According to the humanized matrix extraction process, it is mainly divided into four parts ( figure 1 ), first shred the placental tissue, then remove blood protein and cell protein, extract extracellular matrix protein components again, and finally sterilize and dialyze into TBS solution.

[0047] 1.1 Prepare reagents

[0048] (1) 2M Tris buffer, pH...

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Abstract

The invention relates to a novel animal source-free and feed layer-free human pluripotent stem cell culture system. Extracellular matrix protein, namely a humanized matrix, which can support long-term culture of human embryonic stem cells and is extracted from placenta by a pepsase digestion and urea extraction method, a component prepared from human plasma by a NaCl precipitation method is prepared into a humanized medium, and the humanized medium and the humanized matrix form the animal source-free and feed layer-free culture system together. The culture system can maintain the potentialityof self-renewal and differentiation of human embryonic stem cells for a long term, and an animal source-free human induced pluripotent stem cell clone can be established in the culture system, so theanimal source-free and feed layer-free culture system which is low in cost and suitable for amplification culture lays a foundation for clinical application of pluripotent stem cells.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a novel human pluripotent stem cell culture system without animal source and feeder layer, which is suitable for long-term culture of human pluripotent stem cells. Background technique [0002] The typical characteristic of human pluripotent stem cells is that they continuously self-renew in vitro while maintaining the potential to differentiate into cells of various germ layers. Because the establishment of human embryonic stem cells involves ethical and legal issues, its research and application are subject to certain restrictions. Induced pluripotent stem cells have solved the ethical controversy of human embryonic stem cells well. At the same time, because they have similar characteristics to human embryonic stem cells, that is, self-renewal and differentiation potential, they have brought great potential to regenerative medicine, especially personalized treatment. hope...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/074C12N5/0735
Inventor 马跃王奇慧牟晓宁
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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