Method for improving reprogramming efficiency of somatic cells

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

Active Publication Date: 2010-03-10
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: B6D2F1 female mice (C57BL / 6·DBA / 2) and ICR female mice aged 8 to 12 weeks provided oocytes and granulosa cells respectively, and B6D2F1 female mice and ICR female mice were also used as recipients of embryo transfer .

[0016] 2. Preparation of oocytes and granulosa cells: B6D2F1 (C57BL / 6 DBA / 2) and ICR female mice were injected intraperitoneally with 10 IU pregnant horse serum gonadotropin (PMSG), and 10 IU human chorion was injected 48 hours after superovulation Gonadotropin (hCG), 15 hours after the injection of hCG, the mice were killed by neck dislocation, the oviducts were taken, and the enlarged part was stripped under a microscope to obtain cumulus-oocyte complexes (COCs). COCs were treated with 300U / ml hyaluronidase to separate granulosa cells from oocytes. Oocytes were washed twice with HEPES-CZB culture medium and put into CZB culture medium. At 37°C with 5% CO 2 cultured under air and saturated humidity condit...

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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 maturedevelopmental 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 invention relates to a method for improving the reprogramming efficiency of somatic cells. Background technique [0002] Somatic cell nuclear transfer technology (SCNT) has broad application prospects in agriculture and regenerative medicine. In agriculture, it can be used for multiplication of improved varieties and protection of endangered animals, and it can be combined with embryonic stem cell technology for therapeutic cloning (Xiangzhong Yang et al., 2007). However, the low in vitro and in vivo development rate of somatic cell cloning technology seriously hinders its application. In order to improve the cloning efficiency, people have tried many methods to improve the cloning efficiency. The main research directions include the activation timing of the reconstituted embryo and the influence of DMSO ( T.Wakayama et al., 2001), effects of cytokinesis inhibitors, effects of enucleation and injection timing (S.Wakayama et al., 2003), electrofusi...

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Application Information

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Patent Type & Authority Applications(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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