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Novel method for therapeutic cloning with substitution of oocyte by developing embryo after cleavage

A technology for embryonic stem cells and embryos, applied in the field of obtaining mouse nuclear transfer recipients, can solve problems such as limited sources and recipient-oocyte limitations

Inactive Publication Date: 2012-11-21
赵巍
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as a receptor for reprogramming in cloning technology, the acquisition of oocytes is subject to many restrictions, and at the same time faces medical and ethical issues
Using fertilized eggs as recipients opens up a new avenue for therapeutic cloning, but sources remain limited

Method used

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  • Novel method for therapeutic cloning with substitution of oocyte by developing embryo after cleavage
  • Novel method for therapeutic cloning with substitution of oocyte by developing embryo after cleavage
  • Novel method for therapeutic cloning with substitution of oocyte by developing embryo after cleavage

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Experimental program
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Embodiment Construction

[0021] Materials and Methods

[0022] embryo culture

[0023] Every 20-30 embryos were placed in 30 μl of KSOM culture medium (CHEMICON) drops, covered with mineral oil, and incubated at 37°C, 5% CO 2 cultured in an incubator. All room temperature operations were performed with a modified Hepes-buffered CZB medium (Chatot et al., 1990) instead of KSOM. Normal temperature operation refers to the operation performed outside the incubator.

[0024] Recipient Embryo Production and Metaphase Synchronization

[0025]B6D2F1 (C57BL / 6×DBA / 2) female mice aged 6-8 weeks were intraperitoneally injected with 10IU PMSG (pregnant horse serum gonadotropin), and 48 hours later injected with 10IU hCG (human chorionic gonadotropin) and mixed with B6D2F1 male mice mating. The post-cleavage-developed 2-cell stage embryos were cultured in KSOM culture droplets from the oviducts of fertilized female mice. The blastomeres were fused according to the time of hCG injection of pregnant mice. The ...

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Abstract

The invention relates to a method for obtaining a mice's nuclear transfer receptor and is characterized in that a developing embryo after cleavage is adopted as the nuclear transfer receptor. The nuclear transfer receptor is obtained through the following steps of fusing blastomeres of the developing embryo after cleavage as a single-cell embryo, and synchronizing the single-cell embryo into the metaphase of mitosis. The invention also relates to a method for preparing an embryonic stem cell with the mice's nuclear transfer receptor and a method for obtaining a cloning mice with the mice's nuclear transfer receptor. Moreover, the invention solves the problem of insufficient reprogramming receptors to a large extent, and can provide a novel concept of the selection of nuclear transfer receptors for therapeutic cloning.

Description

technical field [0001] The invention relates to a method for obtaining a mouse nuclear transfer recipient, a method for obtaining a mouse embryonic stem cell by using the mouse nuclear transfer recipient, and a method for obtaining a cloned mouse by using the mouse nuclear transfer recipient. The present invention also relates to a new method of therapeutic cloning, in which the developing embryo after cleavage replaces the oocyte as the recipient of nuclear transfer. Background technique [0002] With the deterioration of the environment, the extension of human age, the abuse of pesticides and antibiotics, there are more and more difficult human diseases. Embryonic stem cells (ES cells) provide many opportunities to solve the above problems. Embryonic stem cells can proliferate infinitely in vitro, Under certain conditions, it can induce differentiation in a certain direction, which provides attractive application prospects and huge commercial potential for treating and rep...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/06C12N5/16C12N15/00C12N5/10A01K67/027
Inventor 周琪阿姆贾德·里亚兹赵小阳李伟于洋王柳
Owner 赵巍
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