DIRECTED DIFFERENTIATION AND CO-CULTURE OF HUMAN EMBRYONIC STEM CELL DERIVED OSTEOPROGENITORS AND ENDOTHELIAL CELLS FOR BONE TISSUE ENGINEERING

Abstract

This study aimed to develop efficient protocols for derivation of hESC-OP and hESC-EC with minimal use of xenogenic products. Under feeder-free and serum-free conditions, hESC were differentiated to hESC-OP through stages encompassing embryoid bodies and embryoid body outgrowth. The differentiation of hESC to hESC-OP was confirmed at transcript and protein levels, and differentiation towards osteogenic lineage under osteogenic conditions. Similarly, hESC were differentiated towards hESC-EC under feeder-free and serum-free conditions, using sequential modulation of Wnt, FGF, BMP and VEGF signaling pathways. The characteristics of hESC-EC were confirmed at transcript and protein levels and tube formation assays. Further, towards developing strategies for in vitro prevascularization, we investigated the in vitro functionality of these hESC-OP and hESC-EC using 2-dimensional (2d) and 3-dimensional (3d) co-culture systems. This is the first instance of successfully using hESC-OP as supporting cells for EC differentiation.