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人胎盘来源的间充质祖细胞支持脐带血CD34+细胞中长期培养起始细胞的培养扩增。

Human placenta-derived mesenchymal progenitor cells support culture expansion of long-term culture-initiating cells from cord blood CD34+ cells.

作者信息

Zhang Yi, Li Changdong, Jiang Xiaoxia, Zhang Shuangxi, Wu Ying, Liu Bing, Tang Peihsien, Mao Ning

机构信息

Department of Cell Biology, Institute of Basic Medical Sciences, Beijing, People's Republic of China.

出版信息

Exp Hematol. 2004 Jul;32(7):657-64. doi: 10.1016/j.exphem.2004.04.001.

Abstract

OBJECTIVE

Allogeneic transplantation with umbilical cord blood (UCB) in adult recipients is limited mainly by a low CD34+ cell dose. To overcome this shortcoming, human placenta as a novel source of human mesenchymal progenitor cell (MPC) was incorporated in an attempt to expand CD34+ cells from UCB in vitro.

MATERIALS AND METHODS

Human placenta MPC was isolated and characterized by morphologic, immunophenotypical, and functional analysis. UCB CD34+ cells were expanded by coculture with placental MPC. Suitable aliquots of cells were used to monitor cell production, clonogenic activity, and long-term culture-initiating culture (LTC-IC) output. Finally, the immunoregulatory effect of placental MPC was evaluated by T-cell proliferation assay.

RESULTS

In its undifferentiated state, placental MPC displayed fibroblastoid morphology; was CD73, CD105, CD29, CD44, HLA-ABC, and CD166 positive; produced fibronectin, laminin, and vimentin; but was negative for CD14, CD31, CD34, CD45, HLA-DR, and alpha-smooth muscle actin. Functionally, it could be induced into adipocytes, osteocytes, and chondrocytes. In vitro expansion of UCB hematopoietic cells, when cocultured with placental MPC in the presence of cytokines, was significantly enhanced: CD34+ cells by 14.89 +/- 2.32 fold; colony-forming cell (CFC) by 36.73 +/- 5.79 fold; and LTC-IC by 7.43 +/- 2.66 fold. Moreover, placental MPC could suppress T-cell proliferation induced by cellular stimuli.

CONCLUSION

These results strongly suggest that human placental MPC may be a suitable feeder layer for expansion of hematopoietic progenitors from UCB in vitro.

摘要

目的

成年受体的脐带血(UCB)同种异体移植主要受限于低CD34+细胞剂量。为克服这一缺点,将人胎盘作为人骨髓间充质祖细胞(MPC)的新来源,试图在体外扩增UCB中的CD34+细胞。

材料与方法

通过形态学、免疫表型和功能分析对人胎盘MPC进行分离和鉴定。通过与胎盘MPC共培养来扩增UCB CD34+细胞。使用合适的细胞等分试样监测细胞产量、集落形成活性和长期培养起始培养(LTC-IC)产量。最后,通过T细胞增殖试验评估胎盘MPC的免疫调节作用。

结果

在未分化状态下,胎盘MPC呈成纤维细胞样形态;CD73、CD105、CD29、CD44、HLA-ABC和CD166呈阳性;产生纤连蛋白、层粘连蛋白和波形蛋白;但CD14、CD31、CD34、CD45、HLA-DR和α-平滑肌肌动蛋白呈阴性。在功能上,它可以被诱导分化为脂肪细胞、骨细胞和软骨细胞。在细胞因子存在的情况下,与胎盘MPC共培养时,UCB造血细胞的体外扩增显著增强:CD34+细胞增加14.89±2.32倍;集落形成细胞(CFC)增加36.73±5.79倍;LTC-IC增加7.43±2.66倍。此外,胎盘MPC可以抑制细胞刺激诱导的T细胞增殖。

结论

这些结果强烈表明,人胎盘MPC可能是体外扩增UCB造血祖细胞的合适饲养层。

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