Abstract 13608: Enhanced Wound Healing by Human Pluripotent Stem Cell-Derived Lymphatic Endothelial Lineage Cells
Background: A very common, but not a well investigated lymphatic-related disorder is skin wound. Despite increasing evidence for the importance of lymphatic vessel in wound healing, no studies are available regarding the effects of lymphatic endothelial cells (LECs) differentiated from human pluripotent stem cells (hPSCs) on wound repair. Here, we developed a new protocol to differentiate hPSCs which include human ESCs and iPSCs into LECs and investigated their therapeutic potential of hPSC-LECs on wound healing.
Methods and Results: hPSCs (H1, H9, BJ1-hiPSC) were induced to form embryoid bodies for 7 days and the cells were reseeded onto OP9 cells with VEGF-A, VEGF-C and EGF for 14 days. These cells expressed LEC markers, PROX1 (53.5±6.9%), LYVE-1 (64.1±5.1%), VEGFR-3 (10.5±1.7%) and PODOPLANIN (59.0±7.2%) examined by FACS analysis (percent in parentheses), real-time RT-PCR and immunocytochemistry. A subpopulation of these cells expressed triple and quadruple LEC markers by FACS, indicating successful generation of LECs. Next, from this culture, PODOPLANIN+LYVE1+ (POD/LYVE1+) cells (1 × 105) were isolated by cell sorting at day 14 and injected into the margins of the wound which was created on the backs of nude mice. During the two-week follow-up, the wound size was more decreased in POD+LYVE1+ cell injected mice compared to the PBS-injected mice (31.6±11.5mm2 vs 128.5±36.6mm2, P < 0.01). Immunohistochemical analysis showed that the density of lymphatic vessels stained for LYVE-1 was 2.6-fold higher in the POD/LYVE1+ injected mice compared to the LEC-control. The red fluorescent dye, DiI-labeled injected POD/LYVE1+ cells were colocalized with lymphatic vessels suggesting de novo lymphvasculogenesis. The cell injected group also expressed higher levels of lymphangiogenic cytokines, such as VEGFA, VEGFC, VEGFD, and IGF1, ANG1, HGF, and bFGF compared to the control-injected groups suggesting potent lymphangiogenic activities..
Conclusion: These results indicate that hPSC-derived POD/LYVE-1+ cells can enhance wound healing by their lymphvasculogenic and paracrine roles. This is the first report that hPSC-derived LECs have therapeutic potential in wound repair.
- © 2012 by American Heart Association, Inc.