Generation of functional platelets from human embryonic stem cells in vitro via ES-sacs, VEGF-promoted structures that concentrate hematopoietic progenitors
Generation of functional platelets from human embryonic stem cells in vitro via ES-sacs, VEGF-promoted structures that concentrate hematopoietic progenitors
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DOI：
10.1182/blood-2007-10-117622
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发表时间：
2008-06-01
期刊：
影响因子：
20.3
通讯作者：
Nakauchi, Hirornitsu
中科院分区：
文献类型：
--
作者：
Takayama, Naoya;Nishikii, Hidekazu;Nakauchi, Hirornitsu
人胚胎干细胞（hESCs）可能是输血治疗的替代来源，也是研究造血个体发生的有前途的工具。当我们在C3H10T1/2或OP-9细胞上培养hESCs以促进造血时，我们发现外源性血管内皮生长因子促进了囊状结构的出现，我们将其命名为胚胎干细胞衍生囊（ES-sacs）。这些ES-sacs由多个包囊组成，包囊由细胞单层划分，保留了内皮细胞的一些特性。ES-sacs内的球形细胞主要表达CD34，以及VE-cadherin、CD31、CD41a和CD45，能够在半固体培养中形成造血集落，并在血小板生成素存在下在第24天分化为成熟的巨核细胞。显然，ES-sacs为造血祖细胞提供了一个合适的环境。ES-sacs内的造血祖细胞可以诱导相对大量的成熟巨核细胞，这些细胞随后能够释放血小板，这些血小板在ADP或凝血酶的作用下表现出整合素α II β 3的激活和扩散。因此，这种新的方案提供了一种从hESCs中产生血小板的方法，这可以作为临床输血和血小板生成研究中有效生产血小板的基础。
Human embryonic stem cells (hESCs) could potentially represent an alternative source for blood transfusion therapies and a promising tool for studying the ontogeny of hematopoiesis. When we cultured hESCs on either C3H10T1/2 or OP-9 cells to facilitate hematopoiesis, we found that exogenous administration of vascular endothelial growth factor promoted the emergence of sac-like structures, which we named embryonic stem cell-derived sacs (ES-sacs). These ES-sacs consisted of multiple cysts demarcated by cellular monolayers that retained some of the properties of endothelial cells. The spherical cells inside ES-sacs expressed primarily CD34, along with VE-cadherin, CD31, CD41a, and CD45, and were able to form hematopoietic colonies in semisolid culture and to differentiate into mature megakaryocytes by day 24 in the presence of thrombopoietin. Apparently, ES-sacs provide a suitable environment for hematopoetic progenitors. Relatively large numbers of mature megakaryocytes could be induced from the hematopoietic progenitors within ES-sacs, which were then able to release platelets that displayed integrin alpha II beta 3 activation and spreading in response to ADP or thrombin. This novel protocol thus provides a means of generating platelets from hESCs, which could serve as the basis for efficient production of platelets for clinical transfusion and studies of thrombopoiesis.