Human chorion-derived stem cells: changes in stem cell properties during serial passage
Human chorion-derived stem cells: changes in stem cell properties during serial passage
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DOI：
10.3109/14653249.2010.549121
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发表时间：
2011-05-01
期刊：
影响因子：
4.5
通讯作者：
Hayati, Abdul-Rahman
中科院分区：
文献类型：
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作者：
Fariha, Mohd-Manzor Nur;Chua, Kien-Hui;Hayati, Abdul-Rahman
背景目标。来自人类胎盘组织的胎膜被描述为干细胞的潜在来源。尽管有大量关于羊膜干细胞的文献，但对绒毛膜来源干细胞的干细胞特性的研究有限。方法.主要目的是确定连续传代的人绒毛膜来源的干细胞（hCDSC）的干特性。进行定量聚合酶链反应（PCR）以揭示连续传代的hCDSC中的以下干性基因表达：Oct-4、Sox-2、FGF-4、雷克斯-1、TERT、Nanog（3）、巢蛋白、FZD-9、ABCG-2和BST-1。从传代（P）1至P5评价细胞生长速率。测定P3、P5细胞集落形成单位成纤维细胞（CFU-F）频率及P5细胞多向分化潜能。采用表面标志物CD 9、CD 31、CD 34、CD 44、CD 45、CD 73、CD 90、CD 117、HLA-ABC和HLA-DR、-DP和-DQ比较hCDSC的免疫表型。对P0、P1、P3和P5细胞进行滋养细胞标记物的免疫染色以检测培养中滋养细胞的污染，同时对P5细胞进行细胞遗传学分析以筛选染色体异常。结果与P3和P0相比，hCDSC中间充质谱系的表面标志物在P5时表达更高，表明这些干细胞在连续传代后纯度增加。事实上，除了TERT之外的所有干性基因在P1、P3和P5 hCDSC中表达。此外，人绒毛膜含有高克隆形成前体，CFU-F频率为1：30。成功的成脂、成软骨和成骨分化证明了hCDSC的多谱系潜能。核型分析显示hCDSC连续传代后染色体保持稳定。结论. hCDSC即使在以后的传代中也保持多能潜能，因此是未来细胞治疗的有希望的来源。
Background aims. Fetal membrane from human placenta tissue has been described as a potential source of stem cells. Despite abundant literature on amnion stem cells, there are limited studies on the stem cell properties of chorion-derived stem cells. Methods. The main aim was to determine the stemness properties of serial-passaged human chorion-derived stem cells (hCDSC). Quantitative polymerase chain reaction (PCR) was performed to reveal the following stemness gene expression in serial-passaged hCDSC: Oct-4, Sox-2, FGF-4, Rex-1, TERT, Nanog (3), Nestin, FZD-9, ABCG-2 and BST-1. Cell growth rate was evaluated from passage (P) 1 until P5. The colony-forming unit-fibroblast (CFU-F) frequency of P3 and P5 cells and multilineage differentiation potential of P5 cells were determined. The immunophenotype of hCDSC was compared using the surface markers CD9, CD31, CD34, CD44, CD45, CD73, CD90, CD117, HLA-ABC and HLA-DR, -DP and -DQ. Immunostaining for trophoblast markers was done on P0, P1, P3 and P5 cells to detect the contamination of trophoblasts in culture, while chromosomal abnormality was screened by cytogenetic analysis of P5 cells. Results. The surface markers for mesenchymal lineage in hCDSC were more highly expressed at P5 compared with P3 and P0, indicating the increased purity of these stem cells after serial passage. Indeed, all the stemness genes except TERT were expressed at P1, P3 and P5 hCDSC. Furthermore, human chorion contained high clonogenic precursors with a 1:30 CFU-F frequency. Successful adipogenic, chondrogenic and osteogenic differentiation demonstrated the multilineage potential of hCDSC. The karyotyping analysis showed hCDSC maintained chromosomal stability after serial passage. Conclusions. hCDSC retain multipotent potential even at later passages, hence are a promising source for cell therapy in the future.