Directed Differentiation of Hemogenic Endothelial Cells from Human Pluripotent Stem Cells.
Directed Differentiation of Hemogenic Endothelial Cells from Human Pluripotent Stem Cells.
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DOI：
10.3791/62391
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发表时间：
2021-03-31
期刊：
影响因子：
--
通讯作者：
Hirschi KK
中科院分区：
文献类型：
--
作者：
Nelson EA;Qiu J;Chavkin NW;Hirschi KK
血管广泛分布于身体的所有组织中，并执行不同的功能。因此，从人多能干细胞中衍生出成熟的血管内皮细胞（其排列在血管腔中）对于许多组织工程和再生应用是至关重要的。在体内，原始内皮细胞来源于中胚层谱系，并被指定为特定亚型，包括动脉、静脉、毛细血管、生血和淋巴。生血内皮细胞特别令人感兴趣，因为在发育过程中，它们产生造血干细胞和祖细胞，然后在整个生命中产生所有的血液谱系。因此，创建一个系统，以产生造血内皮细胞在体外将提供一个机会，研究内皮细胞到造血的转变，并可能导致离体生产的人血液制品和减少对人类供体的依赖。虽然存在几种用于衍生祖细胞和原始内皮细胞的方案，但尚未描述从人干细胞产生充分表征的生血内皮细胞。在这里，提供了一种在大约一周内从人胚胎干细胞衍生造血内皮细胞的方法：用GSK 3 β抑制剂诱导原条细胞分化（CHIR 99021），然后是bFGF介导的中胚层谱系诱导，接着是BMP 4和VEGF-A促进的原始内皮细胞发育，最后由维甲酸诱导造血内皮细胞特化。该方案产生了一个明确定义的生血内皮细胞群体，可用于进一步了解其分子调控和内皮-造血转化，这有可能应用于下游治疗应用。这里介绍的是一个简单的协议，定向分化造血内皮细胞从人类多能干细胞在大约一周。
Blood vessels are ubiquitously distributed within all tissues of the body and perform diverse functions. Thus, derivation of mature vascular endothelial cells, which line blood vessel lumens, from human pluripotent stem cells is crucial for a multitude of tissue engineering and regeneration applications. In vivo, primordial endothelial cells are derived from the mesodermal lineage and are specified toward specific subtypes, including arterial, venous, capillary, hemogenic and lymphatic. Hemogenic endothelial cells are of particular interest because, during development, they give rise to hematopoietic stem and progenitor cells, which then generate all blood lineages throughout life. Thus, creating a system to generate hemogenic endothelial cells in vitro would provide an opportunity to study endothelial-to-hematopoietic transition, and may lead to ex vivo production of human blood products and reduced reliance on human donors. While several protocols exist for the derivation of progenitor and primordial endothelial cells, generation of well-characterized hemogenic endothelial cells from human stem cells has not been described. Here, a method for the derivation of hemogenic endothelial cells from human embryonic stem cells in approximately one week is presented: a differentiation protocol with primitive streak cells formed in response to GSK3β inhibitor (CHIR99021), then mesoderm lineage induction mediated by bFGF, followed by primordial endothelial cell development promoted by BMP4 and VEGF-A, and finally hemogenic endothelial cell specification induced by retinoic acid. This protocol yields a well-defined population of hemogenic endothelial cells that can be used to further understand their molecular regulation and endothelial-to-hematopoietic transition, which has the potential to be applied to downstream therapeutic applications. Presented here is a simple protocol for the directed differentiation of hemogenic endothelial cells from human pluripotent stem cells in approximately one week.
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DOI：
10.1126/science.1247391
发表时间：
2014-08-22
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Fox IJ;Daley GQ;Goldman SA;Huard J;Kamp TJ;Trucco M
通讯作者：
Trucco M
影响因子：
16.6
作者：
Dejana E;Hirschi KK;Simons M
通讯作者：
Simons M
影响因子：
16.6
作者：
Fang JS;Coon BG;Gillis N;Chen Z;Qiu J;Chittenden TW;Burt JM;Schwartz MA;Hirschi KK
通讯作者：
Hirschi KK
影响因子：
4.6
作者：
Liu, Chun;Oikonomopoulos, Angelos;Wu, Joseph C.
通讯作者：
Wu, Joseph C.
影响因子：
5
作者：
Lee, Hyunjae;Chung, Minhwan;Jeon, Noo Li
通讯作者：
Jeon, Noo Li