Enrichment of mouse spermatogonial stem cells by melanoma cell adhesion molecule expression.
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.
Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
Mito
M
Kanatsu-Shinohara
Hiroko
H
Morimoto
Takashi
T
Shinohara
D000818:Animals; D000951:Antigens, Neoplasm / Q000378:metabolism; D051929:CD146 Antigen / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D000071858:Epithelial Cell Adhesion Molecule; D005434:Flow Cytometry; D018189:Immunomagnetic Separation; D007248:Infertility, Male / Q000628:therapy; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D034622:RNA Interference; D034741:RNA, Small Interfering; D011994:Recombinant Proteins / Q000378:metabolism; D013091:Spermatogenesis; D013093:Spermatogonia / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation; D033581:Stem Cell Transplantation / Q000009:adverse effects; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D060245:Tetraspanin 29 / Q000378:metabolism
D000951:Antigens, Neoplasm; D051929:CD146 Antigen; C554870:Cd9 protein, mouse; D015815:Cell Adhesion Molecules; D000071858:Epithelial Cell Adhesion Molecule; C497903:Mcam protein, mouse; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; D060245:Tetraspanin 29