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首页> 外文期刊>Molecular and Cellular Biology >Hematopoietic commitment during embryonic stem cell differentiation in culture.
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Hematopoietic commitment during embryonic stem cell differentiation in culture.

机译：胚胎干细胞在培养过程中的造血作用。

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We report that embryonic stem cells efficiently undergo differentiation in vitro to mesoderm and hematopoietic cells and that this in vitro system recapitulates days 6.5 to 7.5 of mouse hematopoietic development. Embryonic stem cells differentiated as embryoid bodies (EBs) develop erythroid precursors by day 4 of differentiation, and by day 6, more than 85% of EBs contain such cells. A comparative reverse transcriptase-mediated polymerase chain reaction profile of marker genes for primitive endoderm (collagen alpha IV) and mesoderm (Brachyury) indicates that both cell types are present in the developing EBs as well in normal embryos prior to the onset of hematopoiesis. GATA-1, GATA-3, and vav are expressed in both the EBs and embryos just prior to and/or during the early onset of hematopoiesis, indicating that they could play a role in the early stages of hematopoietic development both in vivo and in vitro. The initial stages of hematopoietic development within the EBs occur in the absence of added growth factors and are not significantly influenced by the addition of a broad spectrum of factors, including interleukin-3 (IL-3), IL-1, IL-6, IL-11, erythropoietin, and Kit ligand. At days 10 and 14 of differentiation, EB hematopoiesis is significantly enhanced by the addition of both Kit ligand and IL-11 to the cultures. Kinetic analysis indicates that hematopoietic precursors develop within the EBs in an ordered pattern. Precursors of the primitive erythroid lineage appear first, approximately 24 h before precursors of the macrophage and definitive erythroid lineages. Bipotential neutrophil/macrophage and multilineage precursors appear next, and precursors of the mast cell lineage develop last. The kinetics of precursor development, as well as the growth factor responsiveness of these early cells, is similar to that found in the yolk sac and early fetal liver, indicating that the onset of hematopoiesis within the EBs parallels that found in the embryo.
机译：我们报告胚胎干细胞有效地经历体外分化为中胚层和造血细胞，并且此体外系统概括了小鼠造血发育的6.5至7.5天。分化为胚状体（EB）的胚胎干细胞在分化的第4天和第6天发育出类红细胞前体，其中超过85％的EB包含此类细胞。原始内胚层（胶原蛋白IV）和中胚层（Brachyury）的标记基因的比较逆转录酶介导的聚合酶链反应谱表明，造血之前，这两种细胞类型都存在于发育中的EB和正常胚胎中。 GATA-1，GATA-3和vav在造血细胞的早期和/或早期发作期间在EB和胚胎中表达，表明它们可能在体内和体外的造血发育的早期阶段发挥作用。体外。 EB内造血发育的初始阶段发生在没有添加生长因子的情况下，并且不受多种因子（包括白介素3（IL-3），IL-1，IL-6， IL-11，促红细胞生成素和Kit配体。在分化的第10和14天，通过向培养物中添加Kit配体和IL-11来显着增强EB造血功能。动力学分析表明，造血前体在EB内以有序模式发展。原始红系谱系的前体首先出现，大约在巨噬细胞和确定性红系谱系的前体之前24小时。接下来出现双潜性嗜中性粒细胞/巨噬细胞和多谱系前体，最后发展出肥大细胞谱系的前体。这些早期细胞的前体发育动力学以及生长因子反应性与卵黄囊和早期胎儿肝脏中发现的动力学相似，这表明在EB中发生造血作用与在胚胎中发现的相似。

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