Efficient long-term survival of cell grafts after XXXX with thick viable cardiac tissue entirely from pluripotent stem cells.

@entity35 @entity583 @entity519 @entity65
UNASSIGNED: Poor engraftment of cells after transplantation to the heart is a common and unresolved problem in the cardiac cell therapies. We previously generated cardiovascular cell sheets entirely from pluripotent stem cells with cardiomyocytes, endothelial cells and vascular mural cells. Though sheet transplantation showed a better engraftment and improved cardiac function after @entity583 , stacking limitation (up to 3 sheets) by @entity519 hampered larger structure formation and long-term survival of the grafts. Here we report an efficient method to overcome the stacking limitation. Insertion of gelatin hydrogel microspheres (GHMs) between each @entity65 via appropriate spacing and fluid impregnation with GHMs. Fifteen sheets with GHMs (15-GHM construct; >1   mm thickness) were stacked within several hours and viable after 1 week in vitro. Transplantation of 5-GHM constructs (   2   *   10(6) of total cells) to a @entity35 @entity583 model showed rapid and sustained functional improvements. The grafts were efficiently engrafted as @entity65 cells accompanied by functional capillary networks. Large engrafted cardiac tissues (0.8   mm thickness with 40 cell layers) successfully survived 3 months after TX. We developed an efficient method to generate thicker viable tissue structures and achieve long-term survival of the cell graft to the heart.
