Effect of mesenchymal stem cells transfected with lentiviral vector over-expressing Klotho on stem cell survival in a rat myocardium heart failure model

AIM To observe whether bone-derived mesenchymal stem cells(BMSCs)infected with lentiviral vectors bearing Klotho gene will increase the stem cell survival in a rat myocardium of heart failure.METHODS Rat BMSCs were isolated,cultured,and purified by whole bone marrow adherence method for morphology observations. Lentiviral vectors bearing EGFP-Klotho were constructed. The expression of Klotho mRNA in the BMSCs was detected by RT-PCR. SPF SD rats with heart failure (HF) created by subcutaneous injections of isoproterenol (ISO) were randomly divided into three groups: saline-control HF group, BMSCs with transfected Klotho gene group, and BMSCs with transfected EGFP group. Green fluorescin expression was observed on frozen heart section and expression of Klotho mRNA in the rat myocardium of heart failure was detected by RT-PCR. RESULTS BMSCs were successfully cultured by whole bone marrow adherent method and lentiviral vectors over-expressing Klotho were constructed successfully. BMSCs were infected with lentivirus bearing Klotho and the transfection efficiency was above 70%-80% on day 4 after transfection. Green fluorescin distributed around in myocardial tissues was seen in Klotho-BMSCs group and EGFP-BMSCs group 4 weeks after transfection.RT-PCR detection results showed that Klotho mRNA was expressed in the rat myocardium in each group. The expression level in EGFP-Klotho-BMSCs group was significantly higher than that in EGFP-BMSCs group (P<0.05). CONCLUSION Klotho transfection can increase the survival of BMSCs in rat myocardium of heart failure, possibly by its resistance to apoptosis and improvement of myocardial en