Long-term in vitro expansion of human adipose-derived stem cells showed low risk of tumourigenicity

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

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

In the field of cell-based therapy and regenerative medicine, clinical application is the ultimate goal. However, one major concern is: does in vitro manipulation during culture expansion increases tumourigenicity risk on the prepared cells? Therefore, the aim of this study was to investigate the effect of long-term in vitro expansion on human adipose-derived stem cells (ASCs). The ASCs were harvested from lipo-aspirate samples and cultured until passage 20 (P20), using standard culture procedures. ASCs at P5, P10, P15 and P20 were analysed for morphological changes, DNA damage (Comet assay), tumour suppressor gene expression level (quantitative PCR), p53 mutation, telomerase activity, telomere length determination and in vivo tumourigenicity test. Our data showed that ASCs lost their fibroblastic feature in long-term culture. The population doubling time of ASCs increased with long-term culture especially at P15 and P20. There was an increase in DNA damage at later passages (P15 and P20). No significant changes were observed in both p53 and p21 genes expression throughout the long-term culture. There was also no p53 mutation detected and no significant changes were recorded in the relative telomerase activity (RTA) and mean telomere length (TRF) in ASCs at all passages. In vivo implantation of ASCs at P15 and P20 into the nude mice did not result in tumour formation after 4 months. The data showed that ASCs have low risk of tumourigenicity up to P20, with a total population doubling of 42 times. This indicates that adipose tissue should be a safe source of stem cells for cell-based therapy.

Original languageEnglish
Pages (from-to)67-76
Number of pages10
JournalJournal of Tissue Engineering and Regenerative Medicine
Volume8
Issue number1
DOIs
Publication statusPublished - Jan 2014

Fingerprint

Stem cells
Stem Cells
Cell culture
Telomerase
Telomere
Cell- and Tissue-Based Therapy
Gene expression
DNA Damage
Tumors
DNA
Gene Expression
In Vitro Techniques
Mutation
Regenerative Medicine
Comet Assay
p53 Genes
Tumor Suppressor Genes
Nude Mice
Population
Adipose Tissue

Keywords

  • Biosafety
  • Cell expansion
  • Human adipose-derived stem cells
  • Mutation
  • Telomerase
  • Telomere
  • Tumourigenesis

ASJC Scopus subject areas

  • Biomedical Engineering
  • Medicine (miscellaneous)
  • Biomaterials

Cite this

Long-term in vitro expansion of human adipose-derived stem cells showed low risk of tumourigenicity. / Wan Kamarul Zaman, Wan Safwani; Makpol, Suzana; Sathapan, Somasundaram; Kien Hui, Chua.

In: Journal of Tissue Engineering and Regenerative Medicine, Vol. 8, No. 1, 01.2014, p. 67-76.

Research output: Contribution to journalArticle

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