摘要
目的 :建立精原干细胞的培养方法并对培养细胞的生物学特征进行研究。方法 :用饲养层和无饲养层两种方法培养幼家兔精原干细胞 ,在倒置相差显微镜下观察培养细胞的生长和形态变化 ,并对细胞的糖原、脂质及c-kit受体的细胞化学和免疫细胞化学染色结果进行分析。结果 :成功的分离并培养幼家兔精原干细胞。在培养细胞中精原干细胞为主体细胞 ,并可见少量间质细胞。根据精原干细胞体积大小和形态特点 ,可分为大、小两种类型。PAS染色 ,精原干细胞胞质呈阳性反应 ;免疫细胞化学染色显示 ,体积较小的精原干细胞c -kit受体呈强阳性反应 ,体积较大的大精原干细胞呈弱阳性反应 ;间质细胞PAS染色和c -kit受体染色呈阴性反应 ,而脂质染色呈强阳性反应。精原干细胞培养无论有无饲养层 ,均能呈集落状生长 ,但有饲养层的培养 ,细胞的生长明显优于无饲养层培养。结论 :青春期前的睾丸生精小管是精原干细胞最集中、数量最多 ,且容易获取分离的部位 ;精原干细胞的成功培养为今后重建完整的生精细胞系的治疗性移植和对这类定向干细胞的发育及分化潜能的研究提供了细胞模型。
Objective: To investigate the culture methods of rabbit spermatogonial stem cells (SSCs) and their biological characteristics in vitro. Methods: Rabbit SSCs were cultured in the presence and absence of feeder layer respectively. The development and morphologic changes of cultured cells were observed under transmission electron microscope. Then cytochemical test and immunocytochemical stain analysis were carried out. Results: Rabbit SSCs were isolated and cultured successfully. In the cultured cells, the chief were SSCs, with a small quantity of interstitial cells. According to the shape and dimension of SSCs, they are divided into two types. The cytoplasm of SSCs was positively expressed by PAS staining. Immunocytochemical stain suggested that c-kit receptor of the minor SSCs was strongly positive expression, and c-kit receptor of the bigger SSCs was weakly positive expression. For interstitial cells, PAS staining and c-kit receptor staining were negative expression, and lipid staining was strong positive expression. Rabbit SSCs were colony-forming with or without feeder layer, but the growth of SSCs in the presence of feeder layer is much better. Conclusions: Testis seminiferous tubule in the period of preadolescence is the concentration of SSCs, which is relatively easily obtained. The cultivation of SSCs may provide an ideal cell model for the research of committed stem cells’ development and differentiation, which can be applied in remedial transplantation.
出处
《泸州医学院学报》
2005年第1期13-15,共3页
Journal of Luzhou Medical College
关键词
兔精原干细胞
培养
生物学特征
研究
Spermatogonial stem cells
Culture
Biological characteristics
Research
