The impact of long-term in vitro expansion on the senescence-associated markers of human adipose-derived stem cells

Wan Kamarul Zaman Wan Safwani, Suzana Makpol, Somasundaram Sathapan, Chua Kien Hui

Research output: Contribution to journalArticle

27 Citations (Scopus)

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 senescenceassociated genes, cell cycle distribution and ß-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 ß-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 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.

Original languageEnglish
Pages (from-to)2101-2113
Number of pages13
JournalApplied Biochemistry and Biotechnology
Volume166
Issue number8
DOIs
Publication statusPublished - Apr 2012

Fingerprint

Stem cells
Stem Cells
Cell culture
Galactosidases
Cells
HLA-DQ Antigens
cdc Genes
Critical Pathways
Regenerative Medicine
G2 Phase
Cyclin D1
Growth kinetics
HLA-DR Antigens
Growth
Cell- and Tissue-Based Therapy
Analysis of variance (ANOVA)
In Vitro Techniques
Gene expression
Cell Division
Analysis of Variance

Keywords

  • Adipose-derived stem cells
  • Cell-based therapy
  • Long-term culture
  • Regenerative medicine
  • Senescence

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Biology

Cite this

The impact of long-term in vitro expansion on the senescence-associated markers of human adipose-derived stem cells. / Safwani, Wan Kamarul Zaman Wan; Makpol, Suzana; Sathapan, Somasundaram; Kien Hui, Chua.

In: Applied Biochemistry and Biotechnology, Vol. 166, No. 8, 04.2012, p. 2101-2113.

Research output: Contribution to journalArticle

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