Enrichment of Mouse Spermatogonial Stem Cells by Melanoma Cell Adhesion Molecule Expression
Enrichment of Mouse Spermatogonial Stem Cells by Melanoma Cell Adhesion Molecule Expression
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DOI：
10.1095/biolreprod.112.103861
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发表时间：
2012-12-01
影响因子：
3.6
通讯作者：
Shinohara, Takashi
中科院分区：
文献类型：
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作者：
Kanatsu-Shinohara, Mito;Morimoto, Hiroko;Shinohara, Takashi
精原干细胞（SSCs）是精子发生的基础，但由于其稀缺性，研究一直受到阻碍。虽然隐睾手术通常用于获得丰富的SSC种群，但制作隐睾睾丸耗时长，而且该技术并不适用于许多动物物种。在本研究中，我们利用种系干细胞（即培养的SSCs）筛选了一种新的SSCs表面抗原。生殖细胞移植实验表明，ssc表达黑色素瘤细胞粘附分子（melanoma cell adhesion molecule， MCAM），属于免疫球蛋白超家族，介导不依赖阳离子的粘附。虽然MCAM在GS细胞中的过表达不影响移植后SSC集落形成频率或随后的精子发生，但通过短干扰RNA处理在GS细胞中敲低MCAM可减少集落数量，这表明MCAM在维持SSC潜能中发挥作用。CD9(+)EPCAM（低）MCAM(+)KIT（-）表型的野生型成年睾丸细胞的多参数选择导致SSCs富集561倍。从成熟的成年睾丸中富集SSC的新策略的发展将有助于正常睾丸微环境中SSC的分析。
Spermatogonial stem cells (SSCs) provide the foundation of spermatogenesis, but studies are hampered by their scarcity. Although the cryptorchid operation is often used to obtain an enriched SSC population, making cryptorchid testes is time-consuming and the technique is not applicable to many animal species. In the present study, we screened for a new surface antigen on SSCs using germline stem (GS) cells (i.e., cultured SSCs). Germ cell transplantation experiments showed that SSCs express melanoma cell adhesion molecule (MCAM), which belongs to the immunoglobulin superfamily and mediates cation-independent adhesion. Although MCAM overexpression in GS cells did not influence SSC colony formation frequency or subsequent spermatogenesis after transplantation, MCAM knockdown in GS cells by short-interfering RNA treatment reduced colony numbers, suggesting that MCAM plays a role in sustaining SSC potential. Multiparameter selection of wild-type adult testis cells with a CD9(+)EPCAM(low)MCAM(+)KIT(-) phenotype resulted in a 561-fold enrichment of SSCs. Development of a new strategy for SSC enrichment from mature adult testes will facilitate analyses of SSCs in the normal testicular microenvironment.