Hemangioblastic characteristics of fetal bone marrow-derived Flk1+CD31-CD34- cells
Hemangioblastic characteristics of fetal bone marrow-derived Flk1+CD31-CD34- cells
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DOI：
10.1016/s0301-472x(03)00087-0
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发表时间：
2003-07-01
影响因子：
2.6
通讯作者：
Zhao, RCH
中科院分区：
文献类型：
--
作者：
Guo, H;Fang, BJ;Zhao, RCH
目的：研究从胎儿骨髓中分离出的Flk1(+)CD31(-)CD34(-)细胞是否具有成血管细胞（即内皮细胞和造血细胞的祖细胞）的特征。
材料与方法：利用CD45、GlyA和CD34微磁珠对胎儿骨髓中的单核细胞进行负向分选，然后培养形成细胞集落。选取单个集落进行收集。将培养扩增的细胞分别接种在添加内皮细胞生长因子和造血细胞生长因子的细胞外基质凝胶或半固体培养基上。进行免疫化学染色和逆转录 - 聚合酶链反应以鉴定细胞特征。
结果：单个集落中99%的细胞在传代过程中保持Flk1(+)以及CD31/CD34(-)。在细胞外基质凝胶上，Flk1(+)CD31(-)CD34(-)细胞能够生长为对CD31和血管性血友病因子（vWF）呈阳性的血管结构。在血管结构周围存在圆形的CD34(+)细胞。在血管形成前添加血管生成抑制剂苏拉明时，血管结构的形成被阻断。此外，在造血条件下培养的Flk1(+)CD31(-)CD34(-)细胞能够分化为表达GATA - 1、2以及γ、β珠蛋白基因的造血细胞。将它们重新接种在甲基纤维素培养基中后，形成了典型的红细胞集落。
结论：源自胎儿骨髓的Flk1(+)CD31(-)CD34(-)细胞能够分化为内皮细胞和造血细胞。结果表明，胚胎期后的这些Flk1(+)CD31(-)CD34(-)细胞具有成血管细胞的特征，可能在造血和血管疾病方面具有潜在应用价值。（C）2003国际实验血液学学会。由爱思唯尔公司出版。
Objective. To investigate whether Flk1(+)CD31(-)CD34(-) cells isolated from fetal bone marrow (BM) have characteristics of hemangioblasts, i.e., progenitors of endothelial and hematopoietic cells.Materials and Methods. Mononuclear cells from fetal BM were negatively sorted by CD45, GlyA, and CD34 micromagnetic beads, then cultured to form cell colonies. A single colony was harvested. Culture-expanded cells were seeded on ECM gel or semisolid media supplemented with endothelial and hematopoietic growth factors, respectively. Immunochemistry staining and RT-PCR were performed for cell characterization.Results. 99% of cells from the single colony maintained Flk1(+) and CD31/CD34(-) during passaging. On ECM gel, Flk1(+)CD31(-)CD34(-) cells could grow into vascular structure that was positive for CD31 and vWF. There were round CD34(+) cells around the vascular structure. When angiogenesis inhibitor suramin was added before tube formation, formation of vascular structure was blocked. Additionally, Flk1(+)CD31(-)CD34(-) cells cultured on hematopoietic condition could differentiate into hematopoietic cells which expressed GATA-1, 2, and gamma, beta globin gene. After being replated in methylcellulose medium, they formed typical erythroid colonies.Conclusions. Flk1(+)CD31(-)CD34(-) cells derived from fetal BM could differentiate into endothelial and hematopoietic cells. The results suggested that these Flk1(+)CD31(-)CD34(-) cells after embryo stage bear characteristics of hemangioblast and may have potential application for the hematopoietic and vascular diseases. (C) 2003 International Society for Experimental Hematology. Published by Elsevier Inc.