精原干细胞分离、鉴定、培养及其应用进展
徐远飞1,侯 彬1,2,周继昌1*
(1 深圳市慢性病防治中心,深圳 518020;2 上海体育科学研究所,上海 200030)

摘 要:摘 要:雄性哺乳动物睾丸内持续的精子发生是维持其生殖能力的必备条件。精原干细胞(SSCs) 是精子发生的基础,是永久分化成精子的克隆源,它既可以自我更新维持体内干细胞的数量,又可以增殖分化形成各阶段的生精细胞直至成熟精子。现对SSCs 分离、鉴定与培养等技术及其应用的进展进行综述,发现两步酶消化、差异贴壁、磁珠分选等方法是体外分离与纯化SSCs 的常用方法。特异性标志物的研究一直在推进SSCs 的纯化与鉴定,并仍将是SSCs 技术的研究重点之一。培养基、血清、生长因子和饲养层在SSCs 体外培养中至关重要,但长期培养中血清可引起SSCs 分化的问题有待解决。在基础与临床方面的应用中,SSCs 体外培养系统为研究精子发生过程、遗传学、雄性辅助生殖和细胞再生治疗等开辟了新的道路。

Advances in isolation, identification and culture of spermatogonial stem cells and their applications
XU Yuan-Fei1, HOU Bin1,2, ZHOU Ji-Chang1*
(1 Shenzhen Center for Chronic Disease Control, Shenzhen 518020 , China; 2 Shanghai Research Institute of Sports Science, Shanghai 200030, China)

Abstract: Abstract: The continuous spermatogenesis in testis is essential to maintain the reproductive ability of male mammals, while the spermatogonial stem cells (SSCs) are fundamental for this process as “permanent” sources to form sperm clone. The SSCs can not only renew themselves to maintain the number of stem cells, but also can proliferate into different stages of reproductive cells from spermatogenic cells to mature sperms. By summarizing the current advances in the technology of isolation, identification and culture of SSCs and their applications, the two step enzyme digestion, differential attachment, and magnetic bead sorting were found to be the commonly used methods for the separation and purification of SSCs in vitro. The study of specific surface markers has been improving the purification and identification of SSCs, which will continue to be one of the hot topics in the SSCs-related techniques. Culture medium, serum, growth factor and feeders were important factors in SSCs culture, but in the long-term culture, the SSCs differentiation caused by serum was a problem to be solved. For the applications in the basic and clinic medicine, the systematic SSCs methods paved new ways for the study of spermatogenesis, genetics, assisted male reproduction, and regeneration therapy.

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