Overexpression of hTERT increases stem-like properties and decreases spontaneous differentiation in human mesenchymal stem cell lines.
To overcome loss of stem-like properties and spontaneous differentiation those hinder the expansion and application of human mesenchymal stem cells (hMSCs), we have clonally isolated permanent and stable human MSC lines by ectopic overexpression of primary cell cultures of hMSCs with HPV 16 E6E7 and human telomerase reverse transcriptase (hTERT) genes. These cells were found to have a differentiation potential far beyond the ordinary hMSCs. They expressed trophoectoderm and germline specific markers upon differentiation with BMP4 and retinoic acid, respectively. Furthermore, they displayed higher osteogenic and neural differentiation efficiency than primary hMSCs or hMSCs expressed HPV16 E6E7 alone with a decrease in methylation level as proven by a global CpG island methylation profile analysis. Notably, the demethylated CpG islands were highly associated with development and differentiation associated genes. Principal component analysis further pointed out the expression profile of the cells converged toward embryonic stem cells. These data demonstrate these cells not only are a useful tool for the studies of cell differentiation both for the mesenchymal and neurogenic lineages, but also provide a valuable source of cells for cell therapy studies in animal models of skeletal and neurological disorders.
Tsai
Chih-Chien
CC
Stem Cell Laboratory, Department of Medical Research & Education and Orthopaedics & Traumatology, Veterans General Hospital, Taipei, Taiwan.
Chen
Chun-Li
CL
Liu
Hwa-Chung
HC
Lee
Yi-Ting
YT
Wang
Hsei-Wei
HW
Hou
Lein-Tuan
LT
Hung
Shih-Chieh
SC
D055415:Bone Morphogenetic Protein 4; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation; D006651:Histocytochemistry; D052162:Human papillomavirus 16; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D046228:Microarray Analysis; D025341:Principal Component Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019098:Telomerase / Q000378:metabolism*; D014162:Transfection; D014212:Tretinoin
C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; D014212:Tretinoin; C509186:TERT protein, human; D019098:Telomerase