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中国生物工程杂志

China Biotechnology
China Biotechnology  2014, Vol. 34 Issue (7): 38-43    DOI: 10.13523/j.cb.20140706
    
Research on Rurification of Mouse Spermatogonial Stem Cells Using Magnetic Microbeads
HE Ya-nan1, CHEN Xiao-li1, REN Xiao-xia1, HAO Hai-sheng1, QIN Tong1, ZHAO Xue-ming1, LU Yong-qiang2, WANG Dong1
1. The Key Larboratory for Farm Animal Genetic and Utilization of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
2. Animal Husbandry and Veterinary Station of Beijng, Beijng 100107, China
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Abstract  

Enrichment and purification of spermatogonial stem cells (SSCs) is a prerequisite for genetic modification methods by SSCs. Stem cells antibody CD90.2 was used to enrich and purify mice SSCs through magnetic activated cell sorting(MACS) method, flow cytometry analysis(FACS) and Real-Time Polymerase Chain Reaction (RT-PCR) were used to detect the sorting efficiency. The sorting result using FACS showed that the purity of SSCs was 50.11% after sorting. However, the data by RT-PCR suggested that the expression of GATA4, a special-expression gene of sertoli cells, was downregulated about 6 times, and the expression of GFRα-1 and OCT4, two special-expression genes of germ stem cells, was upregulated 6.5 and 5.9 times respectively after-sorting. The relative expression of those three genes indicated that the enrichment efficiency using MACS was 6 times. The deviation between FACS and RT-PCR may come from undissociated magnetic bead and genetically modified fluorescence of SSCs.



Key wordsMagnetic activated cell sorting      Flow cytometric analysis      Real-Time Polymerase Chain Reaction     
Received: 08 April 2014      Published: 25 July 2014
ZTFLH:  Q819  
Cite this article:

HE Ya-nan, CHEN Xiao-li, REN Xiao-xia, HAO Hai-sheng, QIN Tong, ZHAO Xue-ming, LU Yong-qiang, WANG Dong. Research on Rurification of Mouse Spermatogonial Stem Cells Using Magnetic Microbeads. China Biotechnology, 2014, 34(7): 38-43.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20140706     OR     https://manu60.magtech.com.cn/biotech/Y2014/V34/I7/38


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