인간 배아줄기세포의 중배엽 유도를 통한 골, 연골, 심근 분화 향상

Abstract

Human embryonic stem cells (hESCs) derived from the inner cell mass of the blastocyst have self-renewal and had a differentiation potential to differentiate all three germ layer cell which were endoderm, mesoderm and ectoderm. This multilineage differentiation ability of hESCs can be used for regenerative medicine applications. However, hESCs form teratomas, which are tumor-like formations containing three-germ-layer tissues is a major obstacle for the clinical application of hESCs. In this study, we enhanced osteognic, chondrogenic, and cardiomyogenic differentiation of hESCs by mesodermal lineage induction with cytokines, conditioned medium, and biomaterials. We treated bone morphogenetic protein-4 (BMP4) and Fibroblast growth factor (FGF) 2 during the embryoid body (EB) state for mesodermal lineage induction. BMP4 and FGF2 treatment decreased endodermal and ectodermal lineage gene expression and increased mesodermal lineage gene expression. Importantly, the mesodermal lineage-induced EBs underwent enhanced osteogenic and chondrogenic differentiation after differentiation induction. Human mesenchymal stem cell-conditioned medium treatment increased gene expression related to the mesodermal lineage and decreased gene expression related to the endodermal and ectodermal lineage in EBs composed of hESCs. Subsequently osteogenic and chondrogenic differentiation was enhanced compared to untreated EBs. Graphene increased mesodermal lineage, endodermal lineage, and cardiac lineage differentiation. And graphene cultured hESCs showed enhanced extracellular matrix genes expressions. Gold-coated polycaprolactone microsphere increased mesodermal lineage, endodermal lineage, and cardiac lineage differentiation by incorporated to EBs. Enhancement of hESCs differentiation to osteogenic, chondrogenic, and cardiomyogenic by mesodermal lineage induction would advantages for clinical application of hESCs.