Percoll fractionation of adult mouse spermatogonia improves germ cell transplantation.
Percoll fractionation of adult mouse spermatogonia improves germ cell transplantation.
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发表时间：
2004-02
影响因子：
2.9
通讯作者：
Kyu‐Bom Koh;M. Komiyama;Y. Toyama;T. Adachi;C. Mori
中科院分区：
文献类型：
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作者：
Kyu‐Bom Koh;M. Komiyama;Y. Toyama;T. Adachi;C. Mori
目的从成年小鼠睾丸中分离生殖细胞并进行移植。方法采用表达绿色荧光蛋白（GFP）的转基因小鼠作为供体，以区分移植细胞和受体内源性细胞。在10-12周龄时从供体收集生殖细胞，并通过蒸馏分离浓缩精原细胞，并移植到先前在5周龄时用白消安处理以去除内源性生精细胞的受体生精小管中。结果移植后20周，在受体曲细精管中观察到广泛的GFP信号。在14个移植睾丸中有12个（86%）的切片中证实了精子发生和精子的存在。然而，当生殖细胞在没有浓缩的情况下移植时，成功率为零（0/9）。结论成年小鼠睾丸生殖细胞移植到受体曲细精管时，如果细胞群中精原细胞丰富，则可以成功移植，而精原细胞分离法是获得这种细胞群的有效方法。
AIM To isolate and transplant germ cells from adult mouse testes for transplantation. METHODS In order to distinguish transplanted cells from endogenous cells of recipients, donor transgenic mice expressing green fluorescent protein (GFP) were used. Germ cells were collected from the donors at 10-12 weeks of age and spermatogonia were concentrated by percoll fractionation and transplanted into recipient seminiferous tubules that had been previously treated with busulfan at 5 weeks of age to remove the endogenous spermatogenic cells. RESULTS Twenty weeks after the transplantation, a wide spread GFP signal was observed in the recipient seminiferous tubules. The presence of spermatogenesis and spermatozoa was confirmed in sections of 12 out of 14 testes transplanted (86 %). However, when germ cells were transplanted without concentration the success rate was zero (0/9). CONCLUSION Germ cells from adult mouse testes can be successfully transplanted into recipient seminiferous tubules if the cell population is rich in spermatogonia and the percoll fractionation is useful in obtaining such a cell population.