人精原干细胞特异性标志的初步筛选

目的:寻求可能用于人精原干细胞(SSC)分离和纯化的特异性表面标志。方法:通过免疫组化方法,应用造血干细胞(HSC)表面标志c-kit、Thy-1,人胚胎干细胞(ES)表面标志阶段特异性胚胎抗原SSEA-3、SSEA-4、碱性磷酸酶(ALP),原始生殖细胞(PGC)标志SSEA-1以及小鼠SSC表面标志α6和β1整合素对成人及胎儿睾丸SSC的特异性表达进行筛选和鉴定。结果:在成人睾丸组织中,α6整合素在生精小管生殖细胞表面存在较广泛而显著阳性表达,而β1整合素主要在生精小管基底部细胞存在显著阳性染色,Thy-1在成人睾丸生精小管基底部细胞可见散在阳性染色,少量间质细胞中亦可见阳性染色。上述3种抗原标志在成人生殖细胞表面的表达具有一定的特异性。在胎儿睾丸生精小管中,可见SSEA-1在生殖细胞表面存在显著阳性表达,具有明显特异性。结论:α6、β1整合素和Thy-1可作为阳性标志用于人SSC的分选。SSEA-1可作为识别胎儿SSC的特异性标志。

Preliminary Screening of Specific Surface Markers of Human Spermatogonial Stem Cells

OBJECTIVE: To explore the specific surface markers for the isolation and purification of human spermatogonial stem cells (SSC). METHODS: Specific markers of human SSC were screened and identified in fetal and adult testes by immunohistochemical assay, using HSC markers c-kit, Thy-1 and human ES integrins. RESULTS: In human adult testes, the alpha6 integrin extensively and significantly expressed on the surface of most of the germ cells in the seminiferous tubule, and beta1 integrin mainly expressed on the surface of the germ cells residing on or near the basal membrane in the seminiferous tubule. Thy-1 scattering expressed on the surface of some cells of the basal membrane, and on some Leydig cells as well. The three antigen markers expressed on the SSC of human adult testes specifically to some extent. SSEA-1 specifically expressed on the surface of the gonocytes in the fetal testes. CONCLUSION: The alpha6 and beta1 integrins and Thy-1 may be used for the SSC isolation as positive markers. SSEA-1 can be used as an identification marker for the fetus SSC.