Method for generating and inducing pluripotent stem cells by using pig fibroblasts

A technology for pluripotent stem cells and pig fibroblasts, which is applied in the field of pluripotent stem cells, can solve the problems of difficulty in feeding, development into tumors, and complicated nuclear transplantation operations, and achieves the effect of optimizing the iPS induction strategy and facilitating gene manipulation.

Active Publication Date: 2009-12-30
杭州健崃生物科技有限公司
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Problems solved by technology

(3) hES cells are tumorigenic, and may develop into tumors after transplantation into the recipient body. Even if countermeasures such as using SCNT technology and setting suicide genes for transplanted cells may not be able to solve this problem well
However, due to the incomplete reprogramming of the nucleus by nuclear transfer, even if offspring are produced, they are often deformed, and the operation of nuclear transfer is cumbersome and the cycle is long, so the efficiency is low, time-consuming and laborious
In addition, before applying human iPS cells to the clinic, the safety issue, that is, the tumorigenicity issue, needs to be strictly tested
Due to the short lifespan of mice and the difficulty of raising monkeys

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  • Method for generating and inducing pluripotent stem cells by using pig fibroblasts
  • Method for generating and inducing pluripotent stem cells by using pig fibroblasts
  • Method for generating and inducing pluripotent stem cells by using pig fibroblasts

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Embodiment Construction

[0029] In order to make the present invention easier to understand, the present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0030] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology, microbiology, cell biology, immunology and recombinant DNA, which are within the skill of the art. See, e.g., Sambrook, Fritsch, and Maniatis, A Laboratory Guide to Molecular Cloning, 3rd Edition (2002); CURRENT PROTOCOLSIN MOLECULAR BIOLOGY (eds. F.M. Ausubel et al. (1987)); SERIES METHODS INENZYMOLOGY (Academic Press, Inc.): PCR 2: A PRACTICAL APPROACH (M.J.MacPherson, B.D.Hames and G.R.Taylor, (1995)), Harlow and Lane, (1988) ANTIBODIES, A LABORATORY MANUAL and ANIMAL CELLCULTURE (R.I.Freshney, (1987)); W.French Anderson et al....

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Abstract

The invention discloses a method for generating and inducing pluripotent stem cells by using pig fibroblasts. The method comprises the following steps: (1) leading cDNA containing pluripotent stem cell factors into the pig fibroblasts; (2) culturing the pig fibroblasts obtained in the step (1); and (3) authenticating pluripotent stem cell clones. The invention successfully induces cells similar to embryonic stem cells (ES) which have the potential of in vitro differentiation and are easy for genetic manipulation. In the invention, the pig induced pluripotent stem (iPS) cells provide convenience for genetic manipulation and can provide scientific data for further developing differentiation research, building disease models, detecting the security of the iPS cells taking the pig as a model, optimizing the inducing strategy of the iPS cells and accelerating the clinic application of human iPS cells.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to a method for generating induced pluripotent stem cells using pig fibroblasts. Background technique [0002] Stem cells (stem cells) are the initial source of the human body and its various tissue cells, and its most notable biological characteristics are the ability of self-renewal and continuous proliferation, as well as the potential of multidirectional differentiation. Stem cells are divided into somatic stem cells (somatic stem cells) and embryonic stem cells (embryonic stem cells, ES cells) according to different sources. Adult stem cells include those present in adult tissues such as bone marrow mesenchymal stem cells, pancreatic stem cells, and neural stem cells. [0003] In 1981, the isolation and cultivation of ES cells was first successfully done in mice, which is the most widely studied and mature stem cell system so far. Subsequently, the isolation and cultivation of ES ce...

Claims

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Application Information

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IPC IPC(8): C12N15/867C12N5/10
Inventor 裴端卿米盖尔·埃斯特班徐建勇赖良学杨佳银彭梅秀
Owner 杭州健崃生物科技有限公司
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