Abstract 1747: Therapeutic Potential of Mesenchymal Stem Cells Derived from Three Different Layers of Human Fetal Membrane in Hindlimb Ischemia Model

The fetal membrane (FM), which is reported to contain mesenchymal stam cells (MSC), has been regarded as an ideal source for regenerative medicine, although it is normally discarded after birth. Little information is available regarding the biological difference and therapeutic potential of MSC obtained from different layers of FM including amnion and chorion. We assessed the hypothesis that human FM-derived MSC (FM-MSC) might have therapeutic effects on hindlimb ischemia. FM was obtained following cesarean section and vaginal delivery of healthy donor mothers after obtaining informed consent. We mechanically and enzymatically separated human FM into three layers; amnion and inner and outer layers of chorion, and isolated dish-adherent cells from each layer of FM. These FM-derived cells showed similar surface antigen expression and pluripotency compared to bone marrow (BM)-derived MSC. Because cultured inner chorion-derived MSC (IC-MSC) secreted large amounts of growth factors including IGF-1(13.6 ± 0.08 ng/106cells/24hrs, p < 0.001 vs. amnion or outer chorion-derived MSC (AM-MSC or OC-MSC)) and HGF (7.94 ± 0.67 ng/106cells/24hrs, p < 0.001 vs. AM-MSC or OC-MSC) compared to AM-MSC or OC-MSC, we chose to use IC-MSC for further in vivo experiment. Hindlimb ischemia was induced in 6-week-old male NOD/SCID mice, which were divided into two groups: IC-MSC transplanted group (1 × 10 6 cells/50 μl PBS, n = 9), and control group (50 μl PBS, n = 10). IC-MSC was injected into ischemic hindlimb muscle at five different points. Laser Doppler perfusion analyses showed that the ischemic/nonischemic blood flow ratio was significantly increased in IC-MSC group compared to control group (0.468 ± 0.093 vs. 0.227 ± 0.045 respectively, p < 0.01). Transplantation of FM-MSC showed significant neovascularization in murine hindlimb ischemia model. FM-MSC might have a potential to be an ideal source for treatment of ischemic cardiovascular disease.