Transfection of the KCNMA1 Lentiviral into the Mesenchymal Stem Cells in vitro and Its Function Assay

Abstract: Mesenchymal stem cells have a high degree potential of proliferation， self-renewal and multidirectional differentiation. Large conductance calcium-dependent potassium channel， subfamily M， alpha member 1 (KCNMA1) mediated intracellular K+ outflow， so that it made the membrane hyperpolarization and reduced cell excitability. The study through the preparation KCNMA1 recombinant lentivirus and blank vectors lentivirus to transfected into the stem cells， tested the multiplicity of infection (MOI)， detected the expression changes of KCNMA1 and the microenvironment electrolyte concentration before and after KCNMA1 transfected into cells. Results showed that we transfected KCNMA1 and blank vectors lentiviral successfully into the stem cells. The results of RTPCR and Western blot showed that KCNMA1 over-expressed in the transfected KCNMA1 cells compared with the blank vector cells and untransfected cells， and K+ concentration in the microenvironmental of transfected KCNMA1 cells was higher than blank vector cells and untransfected cells. It is confirmed that we had successfully transfected the lentiviral containing KCNMA1 into the BM-MSCs and the cells over-expressed KCNMA1 and performed its functions， laying the foundation for in vivo studies about KCNMA1 and stem cell therapy related diseases.

CSTR: 32200.14.cjcb.2012.11.0006