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Effects of human yolk sac endothelial cells on supporting expansion of hematopoietic stem/progenitor cells from cord blood.

机译:人卵黄囊内皮细胞对支持脐血造血干/祖细胞扩增的影响。

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In order to investigate the effects of human yolk sac-derived endothelial cells (hYSECs) on the expansion of human hematopoietic stem/progenitor cells (HS/PCs) from umbilical cord blood (UCB) in vitro, we purified hYSEC-like cells from 4-5 week human yolk sacs, which were morphologically similar to endothelial cells and expressed CD31, CD144 and vWF characteristics of endothelial cells. Then we isolated CD34(+) cells from UCB in culture under three different conditions: with hematopoietic cytokines (CKs), contact-coculture or noncontact-coculture with hYSECs supplemented with CKs, and found that the contact-coculture system had the strongest expansion efficiency in the total cells' (TCs) ability to form HPP-CFCs. Erythroid burst-forming units (BFU-E) increased 52.35-fold, 20.26-fold and 27.77-fold, respectively, compared with pre-expansion. We detected that the mRNA of Notch ligands such as Jagged1, Delta1 and Delta4 could express in hYSECs after contacted culture with UCB-CD34(+) cells but not the noncontacted cells by RT-PCR analysis. Therefore, we concluded that the contact-coculture system supplemented with CKs could support the expansion of UCB-HS/PCs in vitro, especially high potential proliferative colony-forming cells (HPP-CFC) and BFU-E, perhaps owing to Notch signal pathway.
机译:为了研究人卵黄囊来源的内皮细胞(hYSECs)在体外从脐带血(UCB)扩增人造血干/祖细胞(HS / PCs)的作用,我们从4种纯化的hYSEC样细胞中-5周卵黄囊,在形态上与内皮细胞相似,并表达内皮细胞的CD31,CD144和vWF特征。然后我们在三种不同条件下从培养物中的UCB中分离出CD34(+)细胞:与造血细胞因子(CKs),接触共培养或与CKs互补的hYSEC进行非接触共培养,发现接触共培养系统具有最强的扩增效率总细胞(TC)形成HPP-CFC的能力。与扩展前相比,类红细胞爆发形成单位(BFU-E)分别增长了52.35倍,20.26倍和27.77倍。我们通过RT-PCR分析检测到与UCB-CD34(+)细胞接触培养后,Notch配体如Jagged1,Delta1和Delta4的mRNA可以在hYSECs中表达,而非接触细胞则没有。因此,我们得出结论,补充有CKs的接触共培养系统可以支持UCB-HS / PCs的体外扩增,尤其是高潜能增殖集落形成细胞(HPP-CFC)和BFU-E,这可能是由于Notch信号通路引起的。

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