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Importance of Sox2 in maintenance of cell proliferation and multipotency of mesenchymal stem cells in low-density culture.

Authors
 D. S. Yoon  ;  Y. H. Kim  ;  J. W. Lee  ;  S. Paik  ;  H. S. Jung 
Citation
 Cell Proliferation, Vol.44(5) : 428-440, 2011 
Journal Title
 Cell Proliferation 
ISSN
 0960-7722 
Issue Date
2011
Abstract
OBJECTIVES: This study has aimed to repopulate 'primitive' cells from late-passage mesenchymal stem cells (MSCs) of poor multipotentiality and low cell proliferation rate, by simply altering plating density. MATERIALS AND METHODS: Effects of low density culture compared t high density culture on late-passage bone marrow (BM)-derived MSCs and pluripotency markers of multipotentiality were investigated. Cell proliferation, gene expression, RNA interference and differentiation potential were assayed. RESULTS AND CONCLUSIONS: We repopulated 'primitive' cells by replating late-passage MSCs at low density (17 cells/cm(2) ) regardless of donor age. Repopulated MSCs from low-density culture were smaller cells with spindle shaped morphology compared to MSCs from high-density culture. The latter had enhanced colony-forming ability, proliferation rate, and adipogenic and chondrogenic potential. Strong expression of osteogenic-related genes (Cbfa1, Dlx5, alkaline phosphatase and type Ι collagen) in late-passage MSCs was reduced by replating at low density, whereas expression of three pluripotency markers (Sox2, Nanog and Oct-4), Osterix and Msx2 reverted to levels of early-passage MSCs. Knockdown of Sox2 and Msx2 but not Nanog, using RNA interference, showed significant decrease in colony-forming ability. Specifically, knockdown of Sox2 significantly inhibited multipotentiality and cell proliferation. Our data suggest that plating density should be considered to be a critical factor for enrichment of 'primitive' cells from heterogeneous BM and that replicative senescence and multipotentiality of MSCs during in vitro expansion may be predominantly regulated through Sox2.
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/94165
DOI
10.1111/j.1365-2184.2011.00770.x
Appears in Collections:
1. Journal Papers (연구논문) > 1. College of Medicine (의과대학) > Medical Research Center (임상의학연구센터)
1. Journal Papers (연구논문) > 1. College of Medicine (의과대학) > Dept. of Orthopedic Surgery (정형외과학교실)
1. Journal Papers (연구논문) > 1. College of Medicine (의과대학) > Yonsei Biomedical Research Center (연세의생명연구원)
Yonsei Authors
백승일(Paik, Seung Il) ; 윤동석(Yoon, Dong Suk) ; 이진우(Lee, Jin Woo) ; 정호선(Jung, Ho Sun)
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Full Text
http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2184.2011.00770.x/abstract
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