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Method for improving reprogramming efficiency of somatic cells

A somatic and reprogramming technology, applied in the field of improving the efficiency of somatic cell reprogramming, can solve the problems of only blastocyst development rate, reducing the level of DNA methylation in cells, and not improving in vitro development efficiency.

Active Publication Date: 2011-06-08
INST OF ZOOLOGY CHINESE ACAD OF SCI
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Problems solved by technology

Experiments have shown that 5-aza-dC treatment of donor cells can reduce the DNA methylation level of cells, but the in vitro developmental efficiency after nuclear transfer has not improved
Although SAH treatment of donor cells can significantly increase the in vitro development rate of cloned embryos, the blastocyst development rates of the treatment group and the control group are only 33.8% and 26.1%.

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  • Method for improving reprogramming efficiency of somatic cells

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Embodiment 1

[0014] 1. Materials and methods

[0015] 1. Animals: 8-12-week-old B6D2F1 female mice (C57BL / 6·DBA / 2) and ICR female mice provided oocytes and granulosa cells, respectively, and B6D2F1 female mice and ICR female mice were also used as recipients for embryo transfer. .

[0016] 2. Preparation of oocytes and granulosa cells: B6D2F1 (C57BL / 6·DBA / 2) and ICR female mice were injected intraperitoneally with 10IU pregnant horse serum gonadotropin (PMSG), and 10IU human chorion was injected 48 hours after superovulation. Gonadotropin (hCG), 15h after injection of hCG, the mice were sacrificed by cervical dislocation, the fallopian tubes were removed, and the enlarged part was dissected under a microscope to obtain cumulus-oocyte complexes (COCs). COCs were treated with 300U / ml hyaluronidase to separate granulosa cells and oocytes. Oocytes were washed twice with HEPES-CZB medium and placed in CZB medium. at 37°C with 5% CO 2 The cells were cultured under the conditions of high air a...

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Abstract

The invention provides a method for improving the reprogramming efficiency of somatic cells, a method for improving the in vitro development rate of a cloned embryo, a method for improving the mature developmental rate of the cloned embryo, a method for improving the establishment efficiency of in vitro nuclear transplantation of non-human embryonic stem cell line, and an in vitro processing method for a reconstructed embryo used for embryo transplantation, which all adopt m-carboxycinnamic acid bishydroxamide to treat the reconstructed embryo. The invention also provides application of the m-carboxycinnamic acid bishydroxamide in the preparation of a medicament for improving the reprogramming efficiency of the somatic cells.

Description

technical field [0001] The present invention relates to a method for improving the reprogramming efficiency of somatic cells. Background technique [0002] Somatic cell nuclear transfer (SCNT) technology has broad application prospects in agriculture and regenerative medicine. In agriculture, it can be used for breeding of fine breeds and protection of endangered animals, and combined with embryonic stem cell technology can be used for therapeutic cloning (Xiangzhong Yang et al., 2007). However, the low in vitro and in vivo development rates of somatic cell cloning technology seriously hinder its application. In order to improve the cloning efficiency, many methods have been tried to improve the cloning efficiency. The main research directions include the activation timing of reconstituted embryos and the effect of DMSO ( T. Wakayama et al., 2001), effects of cytokinesis inhibitors, effects of enucleation and injection timing (S. Wakayama et al., 2003), electrofusion (Ogura...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/06A61K31/195
Inventor 周琪王柳代相鹏侯晓军范勇郝捷
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
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