Abstract: Both bone marrow-derived mesenchymal stem cells(MSCs) therapy and gene therapy have been applied to animal studies and clinical trials. The goal of this study was to compare the effects of three types of MSCs transplantation. In the first part of the study, MSCs were transduced with the pEGFP-C1 and transfected with the pEGFP-C1/Akt in a model of bilateral hind limb ischemia in rats. Following gene transduction, the production of Akt and VEGF protein was more in the culture media of GFP-Akt/MSCs group than that in those of the GFP/MSCs group. In the second part of the study, Sprague-Dawley rats(n=10/group) underwent surgery to create bilateral hind limb ischemia and were randomized into two groups consisting of a GFP/Akt-MSCs group(MSCs suspension, 1×10⁷ MSCs/100 μL transfected pEGFP-C1/Akt) and a GFP-MSCs group(MSCs suspension, 1×10⁷ MSCs/100 μL). These PEGFP-C1/Akt and PEGFP-C1 transfected MSCs suspensions were slowly infused into the adductor muscles of the rat’s left hind limb, while the rat’s right hind limb in the control group received an equal volume of PBS. Endpoints includes angiographic analysis, evaluation of capillary density, immunohistochemistry for von Willebrand factor (vWF), immunodetection of Akt and VEGF protein, RT-PCR of VEGF and Akt mRNA levels in vitro and in vivo. Our data indicate that the tissue perfusion can improve capillary density and the mature of vasculature in the GFP-Akt/MSCs group compared to that in the GFP-MSCs group or control group in the rat model of bilateral hind limb ischemia. Transplantation of MSCs transfected with Akt gene may become the future therapy for hind limb ischemia.

Key words: Bone mesenchymal stem cell, pEGFP-C1/Akt, Hindlimb Ischemia, Angiogenesis