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目的研究冻存的大鼠睾丸组织复苏后精原干细胞(SSCs)体外分离培养的生物学特性。方法用二甲基亚砜(DMSO)作低温保护液,于液氮中冻存。4个月后复苏冻存的睾丸组织,进行SSCs的分离纯化,在小鼠胚胎成纤维细胞(MEF)饲养层进行培养,并利用碱性磷酸酶(AKP)鉴定SSCs。结果从冻存睾丸组织分离的精原干细胞在MEF饲养层上的正常形态、增殖能力与新分离的精原干细胞无明显不同。结论大鼠睾丸组织的冷冻复苏并不影响精原干细胞其原有的生物学特性,包括其正常的贴壁、形态和增殖能力。
Objective To study the biological characteristics of isolated and cultured spermatogonial stem cells (SSCs) from frozen rat testes in vitro. Methods DMSO was used as cryogenic protection solution and stored in liquid nitrogen. After 4 months, the frozen testis tissues were resuscitated, SSCs were isolated and purified, and cultured in mouse embryonic fibroblasts (MEF) feeder layer. SSCs were identified by alkaline phosphatase (AKP). Results Normal morphology and proliferation of spermatogonial stem cells isolated from cryopreserved testicular tissue on the MEF feeder layer were not significantly different from that of newly isolated spermatogonial stem cells. Conclusion The cryopreservation of rat testis does not affect the original biological characteristics of spermatogonial stem cells, including its normal adhesion, morphology and proliferation.