A limited temporal window for the derivation of multilineage repopulating hematopoietic progenitors during embryonal stem cell differentiation in vitro
A limited temporal window for the derivation of multilineage repopulating hematopoietic progenitors during embryonal stem cell differentiation in vitro
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DOI：
10.1182/blood.v88.4.1266.bloodjournal8841266
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发表时间：
1996-08-15
期刊：
影响因子：
20.3
通讯作者：
Ansell, JD
中科院分区：
文献类型：
--
作者：
Hole, N;Graham, GJ;Ansell, JD
胚胎干细胞已被证明在体外分化成所有造血谱系。这已被成功地用作研究造血过程中发生的遗传事件的一种方法，然而，由于无法定义胚胎干细胞（ES）在体外产生原始造血干细胞的系统，对中胚层前体向造血谱系的定型的研究受到限制。使用集落形成试验（CFU-A），我们确定了在体外ES细胞分化过程中，在没有外源性条件培养基或基质细胞培养物的情况下，可以检测到原始多系造血前体的最早时间点是4天。从该时间点接受分化的ES细胞的致死辐射的成年受体小鼠存活超过3周;在三个实验中的两个中，这些动物的外周血含有ES衍生的后代。荧光激活细胞分选（FAGS）在移植后12周在淋巴和骨髓区室中发现ES衍生的CD 45（+）造血细胞，提示第4天分化的ES细胞群含有原始造血前体。在这个时间点附近的基因表达的初步RT-PCR分析表明存在非常少的造血细胞。这种方法应证明是有用的遗传控制的承诺和维持造血谱系在体外和体内的研究。(C)1996年，美国血液学会。
Embryonal stem cells have been shown to differentiate in vitro into all hematopoietic lineages. This has been used successfully as one approach to the study of genetic events occurring during haematopoiesis, However, studies on the commitment of mesodermal precursors to the hematopoietic lineage have been limited due to the inability to define a system in which embryonal stem (ES) cells will give rise to primitive hematopoietic stem cells in vitro. Using a colony forming assay (CFU-A), we determined that the earliest time point at which primitive multilineage hematopoietic precursors can be detected during ES cell differentiation in vitro in the absence of exogenous conditioned medium or stromal cell culture is 4 days. Lethally irradiated adult recipient mice that received differentiated ES cells from this time point survived for more than 3 weeks; and in two out three experiments, peripheral blood from these animals contained ES-derived progeny, Fluorescence activated cell sorting (FAGS) found ES-derived CD45(+) hematopoietic cells in both lymphoid and myeloid compartments at 12 weeks posttransplantation, suggesting that the population of day 4 differentiated ES cells contains primitive hematopoietic precursors. A preliminary RT-PCR analysis of gene expression around this time point suggests that there are very few hematopoietic cells present. This approach should prove useful in studies of genetic control of commitment to and maintenance of hematopoietic lineages in vitro and in vivo. (C) 1996 by The American Society of Hematology.