Method for efficiently separating mouse spermatogonial stem cells

A spermatogonial stem cell and mouse technology, applied in the biological field, can solve the problems of increasing the cost of culturing cells, short doubling time, affecting the culture expansion, etc., and achieve the effects of saving the cost of cell culture, improving the separation purity, and the separation method is simple
CN105969723AInactive Publication Date: 2016-09-28INNER MONGOLIA UNIVERSITY
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
INNER MONGOLIA UNIVERSITY
Publication Date
2016-09-28
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a method for efficiently separating mouse spermatogonial stem cells. The method includes the following steps that firstly, mouse testis are collected, and a testicular cell suspension is prepared through a two-step enzyme digestion method; secondly, the testicular cell suspension is inoculated in a porous plate according to a certain density, and a spermatogonial stem cell culture solution is added for in-vitro culture; thirdly, the culture solution is removed, testis cells of different times are cultured through tryptic digestion, and spermatogonial stem cells are sorted from the testis cells through the magnetic activated cell sorting; fourthly, the sorted spermatogonial stem cells are inoculated to an STO trophoblast, and a spermatogonial stem cell culture solution is added for primary culture and subculture of the spermatogonial stem cells. A mouse spermatogonial stem cell system is built through the method including the steps of separation, culture, re-separation and re-culture. The method can achieve the aim of efficiently enriching the spermatogonial stem cells in vitro, and is economical and easy to implement, so that a direction is provided for protection of rare animal varieties and treatment of male infertility.
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Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a method for efficiently isolating mouse spermatogonial stem cells. Background technique

[0002] Spermatogonia stem cells are the source of spermatogenesis in the testis of male mammals. Since 1998, the Brinster laboratory successfully cultured the spermatogonia stem cells in mice for the first time, and proved that the spermatogonia stem cells were separated from the seminiferous tubules. Since it can be cultured in vitro for a long time, many laboratories have also begun to study mammalian spermatogonial stem cells, but the number of spermatogonial stem cells in the mouse testis is small, and the purity of the isolated spermatogonial stem cells is also low, even though it is currently considered that the separation purity is relatively high. The purity of spermatogonial stem cells directly isolated from high THY-1 antibodies is only about 60%, which will affect their culture and e...

Claims

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