Human embryonic stem cell-derived CD34+ cells:: efficient production in the coculture with OP9 stromal cells and analysis of lymphohematopoietic potential
Human embryonic stem cell-derived CD34+ cells:: efficient production in the coculture with OP9 stromal cells and analysis of lymphohematopoietic potential
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DOI：
10.1182/blood-2004-04-1649
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发表时间：
2005-01-15
期刊：
影响因子：
20.3
通讯作者：
Slukvin, II
中科院分区：
文献类型：
--
作者：
Vodyanik, MA;Bork, JA;Slukvin, II
胚胎干细胞（ES细胞）具有作为造血前体细胞移植和造血细胞发育研究的替代来源的潜力。使用人ES（hES）细胞与OP 9骨髓基质细胞的共培养，我们能够获得高达20%的CD 34（+）细胞，并在培养8至9天后从相似数量的初始接种的hES细胞中分离出高达107个纯度超过95%的CD 34（+）细胞。hES细胞来源的CD 34（+）细胞高度富集集落形成细胞，表达造血相关基因加塔-1、加塔-2、SCL/TAL 1和Flk-1的细胞，并且在体外扩增后保留克隆形成潜力。CD 34（+）细胞显示原始造血祖细胞的表型，如CD 90、CD 117和CD 164的共表达所定义，沿着缺乏CD 38表达，并含有醛脱氢酶阳性细胞以及具有维拉帕米敏感性外排罗丹明123能力的细胞。当在存在干细胞因子、Flt 3-L、白细胞介素7（IL-7）的情况下在MS-5基质细胞上培养时。和IL-3，分离的CD 34（+）细胞分化成淋巴样（B和自然杀伤细胞）以及髓样（巨噬细胞和粒细胞）谱系。这些数据表明，通过hES/OP 9共培养产生的CD 34（+）细胞显示出永久造血干细胞的几个特征。
Embryonic stem (ES) cells have the potential to serve as an alternative source of hematopoietic precursors for transplantation and for the study of hernatopoietic cell development. Using coculture of human ES (hES) cells with OP9 bone marrow stromal cells, we were able to obtain up to 20% of CD34(+) cells and isolate up to 107 CD34(+) cells with more than 95% purity from a similar number of initially plated hES cells after 8 to 9 days of culture. The hES cell-derived CD34(+) cells were highly enriched in colony-forming cells, cells expressing hematopoiesis-associated genes GATA-1, GATA-2, SCL/TAL1, and Flk-1, and retained clonogenic potential after in vitro expansion. CD34(+) cells displayed the phenotype of primitive hematopoietic progenitors as defined by co-expression of CD90, CD117, and CD164, along with a lack of CD38 expression and contained aldehyde dehydrogenase-positive cells as well as cells with verapamil-sensitive ability to efflux rhodamine 123. When cultured on MS-5 stromal cells in the presence of stem cell factor, Flt3-L, interleukin 7 (IL-7). and IL-3, isolated CD34(+) cells differentiated into lymphoid (B and natural killer cells) as well as myeloid (macrophages and granulocytes) lineages. These data indicate that CD34(+) cells generated through hES/OP9 coculture display several features of definitive hematopoietic stem cells.