Abstract:
Objective To explore a method of using Caspase-3 small interference ribonucleic acid (siRNA) in vitro to decrease the apoptosis in stable cell lines of bone marrow mesenchymal stem cell (MSC) in order to establish MSC cell lines with stable expression.
Methods Virus particles containing Caspase-3 siRNA were generated in 293FT cells by retroviral packaging system, then were transfected into MSC of rats. And the transfected cells were screened and cultured to get the stable MSC lines. According to the characteristics of retrovirus, the stable expression of the cell lines was identified by Western blot and real-time fluorescent quantitative PCR (RT-QPCR). The apoptosis of stable cell lines and normal MSC were detected by terminal dexynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) and their differences were compared.
Results Compared with the normal cells, the caspase-3 protein expression and mRNA content of stable MSC lines were significantly reduced. In the same condition in vitro, the apoptosis quantity of stable MSC lines significantly decreased compared with the normal cells.
Conclusion Stable cell lines of MSC with stable expression of Caspase-3 siRNA can be obtained by retroviral packaging system. The apoptosis quantity of stable MSC Lines significantly decreased compared with the normal cells.
下载: