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Identifizierung und funktionelle Untersuchung von humanen adulten spermatogonalen und germalen Stammzellen

人类成体精原干细胞和生殖干细胞的鉴定和功能研究

基本信息

批准号：
135740419
负责人：
Professor Dr. Thomas Skutella
金额：
--
依托单位：
Innere Medizin II: Onkologie, Hämatologie, Klinische Immunologie, Rheumatologie und Pulmologie
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2009
资助国家：
德国
起止时间：
2008-12-31 至 2009-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/135740419?language=en
关键词：
Identifizierung und funktionelle Untersuchung von

项目摘要

Our in vitro and in vivo experiments have shown that we are able to generate cultures of human adult pluripotent germline-derived stem cells (haGSCs) from enriched spermatogonia (Conrad et al., 2008). These haGSCs possess expression profiles which resemble those of pluripotent embryonic stem cells (hES). haGSCs can be differentiated in vitro into cell of all three germ layers, and they also form teratomas. These are the first pluripotent stem cells from an adult source which are suitable for use in humans while at the same time being ethically non-controversial. At present, however, only a very few surface markers are available for the prospective isolation of human and/or murine adult spermatogonial stem cells (hSSCs/mSSCs) and haGSCs, including c-kit, Thy-1, α6-integrin, β1-integrin, CD9 and GFRa1, most of which have only been tested in the mouse. In addition, these markers are not highly selective for spermatogonia. Currently employed methods cause substantial loss of cells and enrich not only hSSCs, but other cells involved in spermatogenesis as well. In the proposed project, we plan to develop new methods for the isolation and characterization of highly purified hSSCs and haGSCs, taking advantage of our large library of stem cell-specific antibodies which are promising candidates for a more efficient isolation of these cells. Finally, we will use single cell sorting to establish clones with ES cell-like growth behavior. A central goal is to optimize the culture and selection methods for the generation of hSSCs to obtain sufficient cell numbers from small biopsies for therapeutic applications.

我们的体外和体内实验表明，我们能够从富集的精原细胞中培养出人类多能生殖干细胞 (haGSC)（Conrad 等，2008）。这些 haGSC 具有类似于多能胚胎干细胞 (hES) 的表达谱。 haGSCs可以在体外分化为所有三个胚层的细胞，并且它们还形成畸胎瘤。这些是第一个来自成人的多能干细胞，适用于人类，同时在伦理上没有争议。然而，目前只有极少数表面标记可用于前瞻性分离人类和/或小鼠成体精原干细胞（hSSCs/mSSCs）和haGSCs，包括c-kit、Thy-1、α6-整合素、β1-整合素、CD9和GFRa1，其中大多数仅在小鼠中进行了测试。此外，这些标记物对精原细胞的选择性不高。目前采用的方法会导致细胞大量损失，不仅富集 hSSC，还富集参与精子发生的其他细胞。在拟议的项目中，我们计划利用我们庞大的干细胞特异性抗体库，开发用于分离和表征高度纯化的 hSSC 和 haGSC 的新方法，这些抗体是更有效地分离这些细胞的有希望的候选者。最后，我们将使用单细胞分选来建立具有类似 ES 细胞生长行为的克隆。一个中心目标是优化 hSSC 生成的培养和选择方法，以便从小活检中获得足够的细胞数量用于治疗应用。