Preparation method of animal primary stem cell sterilization culture solution

Abstract

The invention belongs to the technical field of primary stem cell separation and culture, and discloses a preparation method of an animal primary stem cell sterilization culture solution. The preparation method comprises the following steps: under aseptic conditions, taking a pregnant mid-term sheep fetus, shearing corresponding tissues, and standing in a culture solution II; separating by a tissue mechanical separation process, and culturing sheep fetus tissue primary cells in a culture solution I under the conditions of 37 DEG C and 5% CO2 saturated humidity for 48-96 hours; and after the cell clone appears, washing with D-PBS three times, selecting the cells in a favorable state, culturing in a culture solution III for cloning, and culturing to 3-4 generations for cryopreservation for later use. The culture solution can completely ensure no bacterial pollution in the animal primary stem cell separation and culture process within 96 hours. The culture solution can ensure the separated animal primary cells to keep the stem cell characteristics. The culture solution can ensure no bacterial or fungal pollution in the process of separating and culturing the primary stem cells from the 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...

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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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