Rastegar Tayebeh, Habibi Roudkenar Mehryar, Parvari Soraya, Baazm Maryam
Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
Iran J Reprod Med. 2015 Nov;13(11):679-86.
Sertoli cells play a pivotal role in creating microenvironments essential for spermatogonial stem cells (SSCs) self-renewal and commitment to differentiation. Maintenance of SSCs and or induction of in vitro spermiogenesis may provide a therapeutic strategy to treat male infertility.
This study investigated the role of luekemia inhibitory factor (LIF) on the propagation of SSCs and both functions of Sertoli cells on the proliferation and differentiation of these cells.
SSCs were sorted from the testes of adult male mice by magnetic activated cell sorting and thymus cell antigen 1 antibody. On the other hand, isolated Sertoli cells were enriched using lectin coated plates. SSCs were cultured on Sertoli cells for 7 days in the absence or presence of LIF. The effects of these conditions were evaluated by microscopy and expression of meiotic and post meiotic transcripts by reverse transcriptase polymerase chain reaction.
Our data showed that SSCs co-cultured with Sertoli cells in the presence of LIF formed colonies on top of the Sertoli cells. These colonies had alkaline phosphatesase activity and expressed SSCs specific genes. SSCs were enjoyed limited development after the mere removal of LIF, and exhibiting expression of meiotic and postmeiotic transcript and loss of SSCs specific gene expression (p< 0.05).
Our findings represent co-culture of SSCs with Sertoli cells provides conditions that may allow efficient proliferation and differentiation of SSCs for male infertility treatment.
支持细胞在为精原干细胞（SSCs）自我更新和分化创造必要的微环境中发挥关键作用。维持SSCs或诱导体外精子发生可能为治疗男性不育提供一种治疗策略。
本研究调查白血病抑制因子（LIF）对SSCs增殖的作用以及支持细胞对这些细胞增殖和分化的功能。
通过磁激活细胞分选和胸腺细胞抗原1抗体从成年雄性小鼠睾丸中分离SSCs。另一方面，使用凝集素包被的平板富集分离的支持细胞。SSCs在有或无LIF的情况下在支持细胞上培养7天。通过显微镜检查以及逆转录聚合酶链反应检测减数分裂和减数分裂后转录本的表达来评估这些条件的影响。
我们的数据表明，在LIF存在下与支持细胞共培养的SSCs在支持细胞顶部形成集落。这些集落具有碱性磷酸酶活性并表达SSCs特异性基因。仅去除LIF后，SSCs的发育受到限制，表现出减数分裂和减数分裂后转录本的表达以及SSCs特异性基因表达的丧失（p<0.05）。
我们的研究结果表明，SSCs与支持细胞共培养提供了可能允许SSCs有效增殖和分化以治疗男性不育的条件。
