Solution and method for culturing bovine somatic cell cloned embryos

A technology for cloning embryos and culture methods, applied in tissue culture, biochemical equipment and methods, microorganisms, etc., to achieve the effect of increasing the cell rate

Inactive Publication Date: 2010-10-06
NORTHWEST A & F UNIV
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Problems solved by technology

At present, there is no development report on the use of activin A added to the oviduct culture medium to improve the 8 / 16 cell rate, blastocyst rate, hatching rate, etc. of bovine somatic cell cloned embryos. Therefore, a new type of activin A containing The bovine somatic cell clone embryo culture medium is very necessary

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  • Solution and method for culturing bovine somatic cell cloned embryos
  • Solution and method for culturing bovine somatic cell cloned embryos

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Embodiment Construction

[0024] A bovine somatic cell clone embryo culture fluid, comprising commercially available bovine somatic cell clone embryo oviduct culture fluid (mSOF), said bovine somatic cell clone embryo oviduct culture fluid (mSOF) comprising the CaCl of 2mmol / L 2 2H 2 O, 107mmol / L NaCl, 3.2mmol / L KCL, 1.3mmol / L KH 2 PO 4 , 20-80ng / mL of activin A and the rest of the water, practice has proved that the best concentration of activin A is 40ng / mL.

[0025] The activin A is recombinant human activin A or bovine activin A, generally recombinant human activin A, because bovine activin A is generally difficult to obtain.

[0026] A culture method using bovine somatic cell clone embryo culture fluid of the present invention, the culture method is specifically:

[0027] 1) Select a bovine somatic granulosa cell monolayer and heat it at a temperature of 38.5°C with 5% CO 2 And under the culture condition of saturated humidity, cultivate for 3 days;

[0028] 2) From day 4, add activin A at a ...

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Abstract

The invention provides a solution and method for culturing bovine somatic cell cloned embryos. The culture solution comprises merchant oviduct culture solution for bovine somatic cell cloned embryos (mSOF), and comprises activin A as well, wherein the concentration of the activin A is 20 to 80 ng / mL. The culture method comprises: culturing a bovine somatic-cell granular cell monolayer under conventional conditions for 3 days; adding the activin A in a concentration of 20 to 80 ng / mL to the merchant oviduct culture solution for bovine somatic cell cloned embryos (mSOF) from the fourth day; obtaining the solution for culturing bovine somatic cell cloned embryos; and using the solution for culturing bovine somatic cell cloned embryos for 5 days to obtain the bovine somatic cell cloned embryos. The solution and the method have the advantages of improving the 8 / 16 cell rate, blastula rate and hatchability of embryos and significantly raising the gene expression level of Na / K-ATPase, Glut-1, ActR II, Smad2 and the like, and are particularly suitable for application in the field of bovine somatic cell cloning.

Description

technical field [0001] The invention relates to an embryo culture solution and a culture method thereof, in particular to a bovine somatic cell clone embryo culture fluid and a culture method thereof. Background technique [0002] Embryos produced in vitro must be cultured in vitro for a certain period of time, and embryos of good quality can only be transplanted into recipient uterus after selection. Therefore, the in vitro culture of bovine somatic cell cloned embryos is one of the key links in obtaining somatic cell cloned cattle. Embryos developed in vivo have various unknown cytokines secreted by the mother's reproductive tract, and when cultured in vitro, various components must be artificially supplemented. Therefore, only when the cultured environment in vitro is very close to the oviduct and intrauterine environment in vivo can normal development be achieved. Quantity and quality and production of healthy offspring. [0003] The bovine embryo in vitro culture mediu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06A01K67/02
Inventor 华松张涌兰杰刘永刚宋永利王勇胜刘军
Owner NORTHWEST A & F UNIV
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