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Appl Biochem Biotechnol (2012) 166:21012113
DOI 10.1007/s12010-012-9637-4
Wan Kamarul Zaman Wan Safwani & Suzana Makpol & Somasundaram Sathapan & Kien Hui Chua
Received: 18 October 2011 /Accepted: 22 February 2012 / Published online: 7 March 2012# Springer Science+Business Media, LLC 2012
Abstract Human adipose-derived stem cells (ASCs) have generated a great deal of excitement in regenerative medicine. However, their safety and efficacy issue remain a major concern especially after long-term in vitro expansion. The aim of this study was to investigate the fundamental changes of ASCs in long-term culture by studying the morphological feature, growth kinetic, surface marker expressions, expression level of the senescence-associated genes, cell cycle distribution and -galactosidase activity. Human ASCs were harvested from lipoaspirate obtained from 6 patients. All the parameters mentioned above were measured at P5, P10, P15 and P20. Data were subjected to one-way analysis of variance with a Tukey post hoc test to determine significance difference (P<0.05). The data showed that growth of ASCs reduced in long-term culture and the -galactosidase activity was significantly increased at later passage (P20). The morphology of ASCs in long-term culture showed the manifestation of senescent feature at P15 and P20. Significant alteration
W. K. Z. W. Safwani : K. H. Chua
Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia
W. K. Z. W. Safwanie-mail: [email protected]
S. Makpol
Department of Biochemistry, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysiae-mail: [email protected]
S. Sathapan
Subang Jaya Medical Centre, Selangor, Malaysia e-mail: [email protected]
K. H. Chua (*)
Department of Physiology, Faculty of Medicine, Jalan Raja Muda Abdul Aziz, Universiti Kebangsaan Malaysia, 50300 Kuala Lumpur, Malaysiae-mail: [email protected]
Kien. Hui. Chuae-mail: [email protected]
The Impact of Long-Term In Vitro Expansionon the Senescence-Associated Markers of Human Adipose-Derived Stem Cells
2102 Appl Biochem Biotechnol (2012) 166:21012113
in the senescence-associated genes expression levels was observed in MMP1, p21, Rb and Cyclin D1 at P15 and P20. Significant increase in CD45 and HLA DR DQ DP surface marker was observed at P20. While cell cycle analysis showed significant decrease in percentage of ASCs at S and G2/M phase at later passage (P15). Our data showed ASCs cultured beyond P10 favours the senescence pathway and its clinical usage in cell-based therapy may be limited.
Keywords Adipose-derived stem cells . Long-term...