Zhonghua nan ke xue = National journal of andrology · 2007
[Sorting of spermatogonial stem cells in mice].
Аннотация на английскомСтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод в очереди
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Liu Chun-xia, Zhang Ci, Wang Ling-long, Yu Xiao-xiang
Аннотация
OBJECTIVE: To search for an effective method for enriching spermatogonial stem cells in mice.
METHODS: Bilateral artificial cryptorchidism was performed on 20 six-week old male Kunming mice. Three months after the operation, the testes were removed and single cell suspension prepared by two-step enzyme digestion. FITC-conjugated anti-alpha6-integrin antibody and PE-conjugated anti-c-kit antibody were added for adequate time on ice. Then the cells with low side scatter light-scattering properties were sorted and positively stained for alpha6-integrin and negative c-kit expression. And the viability of the isolated cells was assessed by trypan blue exclusion.
RESULTS: The sorted spermatogonial stem cells constitute 2.8% of the testis cells and over 95% of them were viable.
CONCLUSION: FACS can be used to isolate quantities of viable spermatogonial stem cells.
METHODS: Bilateral artificial cryptorchidism was performed on 20 six-week old male Kunming mice. Three months after the operation, the testes were removed and single cell suspension prepared by two-step enzyme digestion. FITC-conjugated anti-alpha6-integrin antibody and PE-conjugated anti-c-kit antibody were added for adequate time on ice. Then the cells with low side scatter light-scattering properties were sorted and positively stained for alpha6-integrin and negative c-kit expression. And the viability of the isolated cells was assessed by trypan blue exclusion.
RESULTS: The sorted spermatogonial stem cells constitute 2.8% of the testis cells and over 95% of them were viable.
CONCLUSION: FACS can be used to isolate quantities of viable spermatogonial stem cells.