A kind of preparation method of animal primary stem cell sterilized culture medium

Abstract

The invention belongs to the technical field of isolating and culturing primary stem cells, and discloses a method for preparing animal primary stem cell sterilized culture fluid. Under sterile conditions, sheep fetuses in the second trimester are taken, and corresponding tissues are cut and left in culture fluid II; Separated by tissue mechanical separation method, with culture medium I at 37 ° C 5% CO 2 Cultivate the primary cells of sheep fetal tissue under saturated humidity conditions for 48h-96h; wash 3 times with D-PBS after the cell clones appear, pick the cell clones in a better state and culture them with culture medium III, and culture them until the 3rd to 4th passage and freeze them for later use . The culture solution configured in the invention can 100% ensure that no bacterial contamination occurs when the animal primary stem cells are isolated and cultured within 96 hours. At the same time, the culture medium used in the present invention can ensure that the isolated animal primary cells maintain the characteristics of stem cells. The invention can ensure that no bacterial or fungal contamination occurs during the process of isolating and culturing primary stem cells from animal tissues.

Description

technical field [0001] The invention belongs to the technical field of isolating and cultivating primary stem cells, and in particular relates to a method for preparing an animal primary stem cell sterilized culture solution. Background technique [0002] At present, animals are an important source and research material of stem cells. Isolation and culture of primary stem cells from animal body tissues is firstly faced with bacterial or fungal contamination from the tissue and during operation, which often leads to the failure of primary stem cell isolation and culture, and the losses caused are sometimes immeasurable, especially for rare wild animals. Animal or high-value animal primary stem cells, the loss is even greater. Usually, when we first isolate primary stem cells from tissues, we add 1% double antibody to PBS and medium to achieve the purpose of sterilization. But in practice, this is far from enough, often leading to contamination of cultured stem cells and fai...

AI Technical Summary

Problems solved by technology

But in practice, this is far from enough, often leading to contamination of cultured stem cells and failure of primary stem cell isolation and culture

[0003] Through the above analysis, the existing problems and defects of the prior art are as follows: in the prior art, primary stem cells are isolated and cultured from animal body tissues, which often leads to contamination of the cultured stem cells, resulting in the failure of primary stem cell isolation and culture

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