Method for separating human spermatogonial stem cells

A technology of spermatogonial stem cells and cells, which is applied in the field of bioengineering, can solve the problems of low purity and few isolated cells, and achieve the effect of high purity and high yield
CN102344909BInactive Publication Date: 2013-09-11RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
Publication Date
2013-09-11
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention belongs to the field of biological engineering and provides a method for separating human spermatogonial stem cells. The method comprises the following steps of: mechanically separating a testicular tissue; processing the testicular tissue into a semi-liquid state; adding digestive enzyme solution I into the liquid testicular tissue in the semi-liquid state; digesting the testicular tissue into a single convoluted tubule; adding digestive enzyme solution II in the convoluted tubule; digesting the convoluted tubule into a single cell; separating reproductive cells from supporting cells in the single cell; separating G protein-coupled receptor 125 (GPR125) positive spermatogonial stem cells from glial cell line-derived neurotrophic factor (GDNF) family receptor alpha 1 (GFRA1) positive spermatogonial stem cells by adopting an immunomagnetic bead cell sorting method; and identifying the separated GPR125 positive spermatogonial stem cells and the GFRA1 positive spermatogonial stem cells by adopting an immune cell chemical method. By the method, a large number of the high-purity human spermatogonial stem cells are obtained, and sufficient cell sources are provided for the research of the human spermatogonial stem cells and the application of the human spermatogonial stem cells in regenerative medicine and reproductive medicine.
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Description

technical field

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

[0002] Stem cells refer to primitive undifferentiated cells that can both self-renew and differentiate into functional daughter cells. At present, stem cell research has become the frontier of life science research at home and abroad, and is of great significance to cell transplantation, tissue engineering and gene therapy in the treatment of human diseases. Spermatogonial stem cells (SSCs) refer to a subtype of type A spermatogonia. Spermatogonial stem cells are the only adult stem cells that pass genetic material to offspring in male animals and humans. Spermatogonia stem cells are also the basis of spermatogenesis in male animals and humans. The spermatogenesis process in mammals is completed in the seminiferous tubules and includes the complex proc...

Claims

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