Immortalization of human precartilaginous stern cells by transfecting SV40Tag / 中华创伤骨科杂志
Chinese Journal of Orthopaedic Trauma
; (12): 251-254, 2009.
Article
em Zh
| WPRIM
| ID: wpr-395823
Biblioteca responsável:
WPRO
ABSTRACT
Objective To establish the strain of immortalized human precartilaginous stem cells (PSCs) which can be a stable cell resource for study of the molecular mechanism of gene targeting on differ-entiation of PSCs. Methods Plasmid pCMVSV40T/PUR containing simian virus 40 large T antigen gene (SV40Tag) was transfected into human PSCs using lipofeetin transfection method. Colonies were isolated by puromycin selection and expanded by many passages. Immunohistochemistry, RT-PCR and Southern blot were used to identify the transfected cells and to detect the expression and integration of SV40Tag in expanded cell lines. Results The positive colonies were isolated and subcultured, named as immortalized precartilaginous stem cells (IPSCs), which were confirmed as positive to fibrnblast growth factor receptor-3 (FGFR-3). The existence of SV40Tag cDNA was detected by Southern blot and the expression of SV40Tag mRNA and protein by RT-PCR and immunohistochemistry. Conclusion IPSCs strain with SV40Tag can be constructed successfully.
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1
Índice:
WPRIM
Tipo de estudo:
Prognostic_studies
Idioma:
Zh
Revista:
Chinese Journal of Orthopaedic Trauma
Ano de publicação:
2009
Tipo de documento:
Article