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Research Article

Human chorion-derived stem cells: changes in stem cell properties during serial passage

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Pages 582-593 | Received 20 May 2010, Accepted 15 Dec 2010, Published online: 13 Jan 2011
 

Abstract

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.

Acknowledgments

The authors are grateful to the Ministry of Sciences, Technology and Innovation Malaysia for sanctioning the project (Sciencefund 02-01-02 SF0288). The authors also wish to thank Dr Srijit Das for editing the article.

Part of the abstract was presented at the 22nd Malaysian Society of Physiology and Pharmacology Scientific Meeting, held at University of Malaya, Kuala Lumpur, Malaysia, from 5.4.2008 to 6.4.2008.

Disclosure of interest: The authors report no conflicts of interest. The authors are responsible for the content and writing of the paper.

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