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人卵黄囊内皮细胞对支持脐血造血干/祖细胞扩增的作用。

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

作者信息

Hu Liangshan, Cheng Lamei, Wang Jian, Zhao Huiping, Duan Huaxin, Lu Guangxiu

机构信息

National Stem Cells Engineering Research Center, Institute of Human Reproductive and Stem Cell Engineering, Central South University, Changsha City, Hunan 410078, China.

出版信息

Cell Biol Int. 2006 Nov;30(11):879-84. doi: 10.1016/j.cellbi.2006.06.006. Epub 2006 Jun 30.

Abstract

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.

摘要

为了研究人卵黄囊来源的内皮细胞(hYSECs)对体外扩增脐带血(UCB)中人类造血干/祖细胞(HS/PCs)的影响,我们从4 - 5周的人卵黄囊中纯化出类hYSEC细胞,这些细胞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的UCB中分离CD34(+)细胞:添加造血细胞因子(CKs)、与添加CKs的hYSECs进行接触共培养或非接触共培养,发现接触共培养系统在总细胞(TCs)形成HPP - CFCs的能力方面具有最强的扩增效率。与扩增前相比,红系爆式集落形成单位(BFU - E)分别增加了52.35倍、20.26倍和27.77倍。通过RT - PCR分析,我们检测到与UCB - CD34(+)细胞接触培养后的hYSECs中能表达Notch配体如Jagged1、Delta1和Delta4的mRNA,而非接触培养的细胞则不能。因此,我们得出结论,添加CKs的接触共培养系统能够在体外支持UCB - HS/PCs的扩增,尤其是高潜能增殖集落形成细胞(HPP - CFC)和BFU - E,这可能归因于Notch信号通路。

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