Chromosomal instability in human mesenchymal stem cells immortalized with human papilloma virus E6, E7, and hTERT genes.
Human mesenchymal stem cells (hMSCs) are expected to be an enormous potential source for future cell therapy, because of their self-renewing divisions and also because of their multiple-lineage differentiation. The finite lifespan of these cells, however, is a hurdle for clinical application. Recently, several hMSC lines have been established by immortalized human telomerase reverse transcriptase gene (hTERT) alone or with hTERT in combination with human papillomavirus type 16 E6/E7 genes (E6/E7) and human proto-oncogene, Bmi-1, but have not so much been characterized their karyotypic stability in detail during extended lifespan under in vitro conditions. In this report, the cells immortalized with the hTERT gene alone exhibited little change in karyotype, whereas the cells immortalized with E6/E7 plus hTERT genes or Bmi-1, E6 plus hTERT genes were unstable regarding chromosome numbers, which altered markedly during prolonged culture. Interestingly, one unique chromosomal alteration was the preferential loss of chromosome 13 in three cell lines, observed by fluorescence in situ hybridization (FISH) and comparative-genomic hybridization (CGH) analysis. The four cell lines all maintained the ability to differentiate into both osteogenic and adipogenic lineages, and two cell lines underwent neuroblastic differentiation. Thus, our results were able to provide a step forward toward fulfilling the need for a sufficient number of cells for new therapeutic applications, and substantiate that these cell lines are a useful model for understanding the mechanisms of chromosomal instability and differentiation of hMSCs.
Takeuchi
Masao
M
Division of Bioresources, National Institute of Biomedical Innovation, Osaka, Japan. takeuchim@nibio.go.jp
Takeuchi
Kikuko
K
Kohara
Arihiro
A
Satoh
Motonobu
M
Shioda
Setsuko
S
Ozawa
Yutaka
Y
Ohtani
Azusa
A
Morita
Keiko
K
Hirano
Takashi
T
Terai
Masanori
M
Umezawa
Akihiro
A
Mizusawa
Hiroshi
H
D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D043171:Chromosomal Instability; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000648:ultrastructure*; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D050725:Papillomavirus E7 Proteins; D000090063:Proto-Oncogene Mas; D012097:Repressor Proteins / Q000235:genetics*; D019098:Telomerase / Q000235:genetics*
C068651:E6 protein, Human papillomavirus type 16; C000715470:MAS1 protein, human; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D000090063:Proto-Oncogene Mas; D012097:Repressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16; C509186:TERT protein, human; D019098:Telomerase