Kit and method for effectively inducing somatic cell phenotype reprogramming

A programming method, a technology for inducing cells, which is applied in the field of efficiently inducing the apparent reprogramming of adult cells, can solve the problems of low induction efficiency, and achieve the effects of short induction period, low research cost, and simple cell reprogramming method

Inactive Publication Date: 2016-06-15
中国人民解放军联勤保障部队第九二〇医院
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] The purpose of the present invention is to solve the problem of low induction efficiency in existing adult cell reprogramming methods, and propose an adult cell apparent reprogramming method for efficiently preparing pluripotent stem cells

Method used

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  • Kit and method for effectively inducing somatic cell phenotype reprogramming
  • Kit and method for effectively inducing somatic cell phenotype reprogramming
  • Kit and method for effectively inducing somatic cell phenotype reprogramming

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Establishment of an efficient method for inducing apparent reprogramming of adult cells.

[0045] In this example, freshwater crucian carp was selected as the preparation material of the inducer, and human skin tissue was used as the experimental material. The skin tissue was derived from the foreskin tissue of adult healthy males. The foreskin tissue was provided by the Urology Department of Kunming General Hospital of Chengdu Military Region. Consent form, samples are limited to use in this study.

[0046] For the basic operation process of efficiently inducing adult cell reprogramming, see figure 1 , including the following steps:

[0047] (1) Preparation of oocyte extract

[0048] The oocyte extract is prepared by repeated freezing and thawing. The specific steps are as follows: select a crucian carp whose yolk is full of oocytes, soak it in 75% alcohol solution for 30 minutes, and beat it on the back of the head after taking it out. Let the crucian carp die, th...

Embodiment 2

[0065] Generation of mouse induced pluripotent stem cells.

[0066] Utilize high-efficiency induced adult cell reprogramming kit for research. Efficient induction of adult cell reprogramming kit contains:

[0067] Inducers of epigenetic reprogramming in adult cells;

[0068] Adult cell and induced cell culture medium.

[0069] In this embodiment, mouse adult cells are used as experimental materials, and the type of adult cells is fibroblasts, and the fibroblasts are derived from mouse skin tissue. The use of experimental animals was approved by the Experimental Animal Use Committee of Kunming General Hospital of Chengdu Military Region.

[0070] Mouse adult cells were prepared according to step (2) in Example 1 for isolation and culture; adult cells were reprogrammed according to step (3) in Example 1; cell characteristics were reprogrammed according to step (4) in Example 1 Identification.

[0071]Pluripotent stem cells are currently an important therapeutic material in ...

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Abstract

The invention discloses a method and kit for effectively inducing somatic cell phenotype reprogramming, and belongs to the field of a cell biology technique. According to the method and the kit, in accordance with problems in an existing somatic cell reprogramming method, a method, which is used for effectively inducing the somatic cell phenotype reprogramming into induced pluripotent stem cells by taking an extract of a natural active material, namely fish oocyte, as an inducing agent, is invented, and the method is applicable to the preparation of the animal pluripotent stem cells. The method and the kit of the invention can be used for effectively inducing the somatic cell phenotype reprogramming into the induced pluripotent stem cells; compared with other methods, the method is high in reprogramming efficiency on somatic cells, and the inducing method is simple, high in safety and relatively low in cost; and the method disclosed by the invention can solve the problem on the source of the pluripotent stem cells, and the method has broad clinical application value and prospect in such fields as regenerative medicine.

Description

technical field [0001] The invention relates to a kit for efficiently inducing apparent reprogramming of adult cells and a method thereof, belonging to the field of cell biology technology. Background technique [0002] Stem cells are a type of cells with the potential of self-renewal and multilineage differentiation. They can be used for the study of the occurrence and development of living organisms, the mechanism of aging and death, and are widely used for regeneration. Medicine (Regenerative medicine) field, etc. At present, the number of pluripotent stem cells isolated from adult tissues is very limited, which often cannot meet the needs of treatment; however, there are serious ethical issues in directly obtaining stem cells from early embryos, and the source of stem cells severely limits its clinical research and development. application. [0003] The emergence of somatic cell reprogramming technology provides new ideas and methods for the source of stem cells. The ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/074C12N5/077C12N5/079C12N5/071
Inventor 潘兴华朱向情陈强
Owner 中国人民解放军联勤保障部队第九二〇医院
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