Long-term Culture of Human SSEA-4 Positive Spermatogonial Stem Cells (SSCs).
Long-term Culture of Human SSEA-4 Positive Spermatogonial Stem Cells (SSCs).
复制标题
DOI：
10.4172/2157-7633.s2-003
复制
发表时间：
2011-11
期刊：
影响因子：
--
通讯作者：
M. Kokkinaki;Ardalan Djourabtchi;N. Golestaneh
中科院分区：
文献类型：
--
作者：
M. Kokkinaki;Ardalan Djourabtchi;N. Golestaneh
最近，我们和其他两个小组已经表明，人精原干细胞（SSCs）有潜力成为多能性在体外培养条件下，并分化成三个胚层的细胞。这一发现可能为退行性疾病的自体细胞治疗开辟新的途径，绕过与人类胚胎干细胞相关的伦理和免疫学问题。此外，人类精原干细胞可用于治疗癌症存活儿童的不孕症。然而，为了将SSC重编程为多能性，或者为了将它们保存用于不育睾丸的再增殖，第一个和限制性的步骤是获得高度纯化的人SSC群体，其可以在体外繁殖和有效培养，保持其分子和细胞特征。尽管各种研究已经尝试鉴定人SSC的分子标记物，但迄今为止，仍然存在与可用于其分离和优化纯化的特定标记物相关的有限信息，所述优化纯化允许分离的人SSC的长期体外培养。在此使用SSEA-4作为分离SSC亚群的最佳标记物，我们显示与用不同标记物分离的其它亚群相比，SSEA-4阳性细胞表达最高水平的SSC基因，并且可以在培养物中维持超过14代，这是我们用其它SSC标记物（包括GPR 125和ITGA 6）无法获得的。此外，我们还建立了一种用于细胞分选和长期培养人SSC-SSEA-4阳性细胞的新技术，可最大限度地提高分选细胞的纯度和活力。我们的研究结果是至关重要的，可用于最有效的分离，纯化和长期培养的精原干细胞在再生医学的临床应用，或用于制备人类精原干细胞自体治疗癌症存活儿童的不孕症。
Recently we and two other groups have shown that human spermatogonial stem cells (SSCs) have the potential to become pluripotent in vitro in defined culture conditions and to differentiate into cells of the three embryonic germ layers. This discovery could open new avenues for autologous cell-based therapy in degenerative diseases, bypassing the ethical and immunological problems related to the human embryonic stem cells. In addition, human SSCs could be used to treat infertility in cancer survival children. However, in order to reprogram SSCs into pluripotency, or to preserve them for repopulation of infertile testes, the first and limiting step is to have access to a highly purified human SSC population that could be multiplied and efficiently cultured in vitro maintaining their molecular and cellular characteristics. Although various studies have attempted to identify molecular markers of human SSCs, to date there is still limited information related to the specific markers that could be used for their isolation and optimized purification that allows long-term in vitro culture of isolated human SSCs. Here using SSEA-4 as an optimal marker for isolation of a subpopulation of SSCs, we show that SSEA-4 positive cells express the highest level of SSC genes compared to other subpopulations isolated with different markers, and can be maintained in culture for over 14 passages which we were unable to obtain with other SSCs markers including GPR125 and ITGA6. In addition, we have established a new technology for cell sorting and long-term culture of human SSC-SSEA-4 positive cells that maximizes the purity and viability of the sorted cells. Our findings are crucial and could be used for the most efficient isolation, purification and long-term culture of SSCs for clinical applications in regenerative medicine, or for preparation of human SSCs for autologous treatment of infertility in cancer survival children.