Development of hematopoietic stem cells from human ES and iPS cells

利用人类 ES 和 iPS 细胞开发造血干细胞

• 批准号: 207511694
• 负责人: Dr. Katharina Hartmann
• 金额: --
• 依托单位: Institute for Stem Cell Biology and Regenerative Medicine
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2012-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/207511694?language=en
• 关键词: Development; hematopoietic; stem; cells; human

The ability to generate hematopoietic stem cells (HSCs) in vitro from embryonic stem (ES) and induced pluripotent stem (iPS) cells would allow replacement of defective hematopoietic cells without the need of human leukocyte antigen (HLA)-matching donors. Several studies have shown that mature hematopoietic cells can be successfully developed in vitro from ES and iPS cells. However, the in vitro generation and maintenance of HSCs, which can reconstitute the whole hematopoietic system in vivo, remains still challenging.The aim of the proposed project is to find optimal culture conditions in order to develop HSCs rather than mature hematopoietic cells in vitro from human ES and iPS cells in a system using stromal cell lines. It has been shown that the microenvironment and the time frame influence the development of HSCs in vitro. Hence, different stromal cell lines and time plans will be applied. Surface marker analyses and subsequent cell sorting of the differentiating cells will allow to enrich HSCs and to deplete undifferentiated cells with teratogenic potential in the culture.Recently, the group of Prof. Weissman identified genes that are differentially expressed in reconstituting HSCs versus non-reconstituting hematopoietic progenitors. Those candidate genes, which may favor the development of HSCs from ES and iPS cells, will be overexpressed using retroviral vectors carrying cDNA. Alternatively, mRNA of the candidate genes will be administered. The influence of these genes will be analyzed by in vitro differentiation and in vivo reconstitution assays. Finally, upon successful hematopoietic reconstitution the ability to tolerate syngenic and allogenic organ transplants will be investigated.

从胚胎干细胞（ES）和诱导多能干细胞（iPS）中体外生成造血干细胞（hsc）的能力将允许替换有缺陷的造血细胞，而不需要人类白细胞抗原（HLA）匹配的供体。一些研究表明，成熟的造血细胞可以在体外成功地从胚胎干细胞和诱导性多能干细胞中培养出来。然而，造血干细胞的体外生成和维持仍然具有挑战性，因为造血干细胞可以在体内重建整个造血系统。该项目的目的是寻找最佳的培养条件，以便在使用基质细胞系的系统中，从人类胚胎干细胞和iPS细胞体外培养出造血干细胞，而不是成熟的造血细胞。已有研究表明，微环境和时间框架影响造血干细胞的体外发育。因此，将采用不同的基质细胞系和时间计划。分化细胞的表面标记分析和随后的细胞分选将允许丰富造血干细胞，并消耗培养中具有致畸潜力的未分化细胞。最近，Weissman教授小组发现了在重建造血干细胞与非重建造血祖细胞中差异表达的基因。这些候选基因可能有利于胚胎干细胞和iPS细胞的发育，它们将在携带cDNA的逆转录病毒载体上过表达。或者，候选基因的mRNA将被管理。这些基因的影响将通过体外分化和体内重组分析来分析。最后，在成功的造血重建后，对同种和异体器官移植的耐受能力将进行研究。