Animal cloned embryo culture solution and method

A technology for embryo culture medium and embryo cloning, applied in biochemical equipment and methods, embryonic cells, culture process, etc., can solve the problem of no animal clone embryo culture medium, so as to improve in vitro development rate and inhibit endoplasmic reticulum stress , the effect of promoting in vitro development

Inactive Publication Date: 2019-04-16
XI AN JIAOTONG UNIV
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Problems solved by technology

[0005] During somatic cell nuclear transfer, operations such as enucleation, fusion, activation, and embryo culture will cause endoplasmic reticulum stress and oxidative stress in animal cloned embryos. At present, there is no in vitro culture medium specifically for animal cloned embryos, especially in In the culture of cloned embryos of gene-edited rabbits and somatic cell nuclear transfer rabbits, it is urgent to propose a culture medium and culture method that can inhibit the endoplasmic reticulum stress of cloned embryogenesis and improve the development rate of animal cloned embryos

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Embodiment Construction

[0025] The present invention will be described in detail below in conjunction with the accompanying drawings and embodiments, which are only used to explain the present invention, rather than limit the protection scope of the present invention.

[0026] 1. Source and preparation of reagents

[0027] Chorionic gonadotropin (HCG) for injection was purchased from Ningbo Sansheng Pharmaceutical Co., Ltd.; sterile water for injection was purchased from Ruicheng Weifu Veterinary Medicine Co., Ltd.; sodium pentobarbital was purchased from Merck. Both DPBS and PBS were purchased from Hyclone Company. PBS-PVA, cytochalasin B, hyaluronidase, trypsin, TUDCA, 6-dimethylaminopurine, M199 culture medium, and DMEM are all sigma products. The electrofusion solution is a Btx product. Fixative, TritonX-100, and Hoechst33342 are all Biyuntian products. FBS is a Gibco product.

[0028] The formula of the embryo culture solution (referred to as A solution) is composed of the following componen...

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Abstract

The invention discloses an animal cloned embryo culture solution and method. The method comprises the steps that tauroursodeoxycholic acid is added into an embryo basal culture solution according to 50-100 microns to obtain the embryo culture solution; animal cloned embryos obtained through nuclear transplantation are transferred into the embryo culture solution for culture. By means of the animalcloned embryo culture solution and method, tauroursodeoxycholic acid is utilized for significantly inhibiting endoplasmic reticulum stress of the animal cloned embryos in the culture stage after thenuclear transplantation, and meanwhile, the in-vitro development rate of the animal cloned embryos is significantly increased. The animal cloned embryo culture solution and method improve the in-vitroculture efficiency of the animal cloned embryos, and have important application value especially in obtaining genetically-modified rabbits and cloned rabbits.

Description

technical field [0001] The invention relates to in vitro embryo culture, in particular to animal cloning embryo culture solution and culture method. Background technique [0002] Nuclear transfer is a cell engineering technique in which a donor cell nucleus is transferred into an enucleated oocyte and a reconstructed embryo is obtained. This technology is of great value in the fields of constructing genetically modified animals, saving endangered species, stem cell therapy, and mitochondrial therapy. Currently, the efficiency of somatic cell nuclear transfer is extremely low. [0003] Reconstructed embryos obtained by nuclear transfer are different from fertilized embryos. The process of constructing embryos involves micromanipulation processes (enucleation, nuclear injection, fusion and activation), and the stimulation of nuclear transfer operations can induce cells to undergo endoplasmic reticulum stress. Excited. Stimulation of in vitro culture conditions can also indu...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073
CPCC12N5/0604C12N2500/30
Inventor 刘恩岐屈鹏祥
Owner XI AN JIAOTONG UNIV
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