Endothelial progenitor cell differentiation using cryopreserved, umbilical cord blood-derived mononuclear cells
Endothelial progenitor cell differentiation using cryopreserved, umbilical cord blood-derived mononuclear cells
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DOI：
10.1111/j.1745-7254.2007.00519.x
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发表时间：
2007-03-01
影响因子：
8.2
通讯作者：
Kim, Hugh C.
中科院分区：
文献类型：
--
作者：
Jang, Jun-ho;Kim, Sun-kyung;Kim, Hugh C.
目的：通过对深低温保存的脐带血单个核细胞(MNC)向内皮祖细胞(EPC)的诱导分化，探讨脐血内皮细胞的分化潜能。方法：将冻存的脐血和外周血的单个核细胞在含内皮细胞生长补充剂的M199培养液中培养14d，用RT-PCR、流式细胞仪和免疫细胞化学方法对内皮细胞进行鉴定。四甲基偶氮唑盐(3-(4，5-dimethylthiazol-2-yl)-2，5-diphenyl-tetrazolium)比色法检测分化内皮祖细胞的增殖情况，ELIS A试剂盒检测血管内皮细胞生长因子浓度。比较了脐带EPC和PB EPC的性能。结果：培养24 h后，大量圆形细胞松散地贴壁于细胞底部，第7天开始出现大量梭形细胞。这些细胞表达Flt-1/VEGFR-1、ecNOS、VE-cadherin、von Willebrand因子等内皮标志物，并分泌血管内皮生长因子。外周血和脐血内皮细胞的内皮标记物图谱无明显差异。增殖和分泌血管内皮细胞生长因子的结果也相似。结论：我们成功地将-196℃保存的脐血细胞培养成具有内皮细胞质量的细胞。这些血管内皮细胞可通过激活邻近的内皮细胞，促进血管生成而用于血管生成治疗。
Aim: To investigate the endothelial differentiation potentiality of umbilical cord blood (UCB), we induced the differentiation of endothelial progenitor cells (EPC) from cryopreserved UCB-derived mononuclear cells (MNC). Methods: MNC from cryopreserved UCB and peripheral blood (PB) were cultured in M199 medium with endothelial cell growth supplements for 14 d. EPC were characterized by RT-PCR, flow cytometry, and immunocytochemistry analysis. The proliferation of differentiated EPC was studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, and vascular endothelial growth factor (VEGF) concentration was measured using an ELIS A kit. Characteristics of UCB-derived EPC were compared with those of PB-derived EPC. Results: A number of round-shaped cells were loosely attached to the bottom after 24 h culture, and numerous spindle-shaped cells began to appear from the round-shaped ones on d 7. Those cells expressed endothelial markers such as, Flt-1/VEGFR-1, ecNOS, VE-cadherin, von Willebrand factor, and secreted VEGF. The patterns of endothelial markers of EPC from PB and UCB did not show striking differences. The results of the proliferation and secretion of VEGF were also similar. Conclusion: We successfully cultured UCB cells stored at -196 degrees C into cells with the quality of endothelial cells. Those EPC could be used for angiogenic therapeutics by activating adjacent endothelial cells and enhancing angiogenesis.