Effective endodermal induction via vitronectin-coated feeder-free culture

Current comprehension of endoderm organ development has enabled researchers to successfully direct pluripotent stem cells (PSCs) to differentiate into endodermal lineage which can give rise to cells of the lungs, liver, pancreas, stomach, and intestine. Nevertheless, variations in the differentiation efficiency have been reported mainly due the utilization of poorly defined matrices in the protocols applied. On this basis, the present study investigated the potential role of recombinant human-Vitronectin-N (VN) in inducing differentiation of human induced pluripotent stem cells (hiPSCs) into endodermal lineage. Differentiation was initiated by applying a previously established sequential step wise endodermal differentiation protocol. Expression of endodermal specific genes characteristic of each differentiation stage was investigated and analyzed by qRT-PCR and immunohistochemistry. Our results demonstrated a significant up-regulation of endoderm related markers characteristic of respective stages of differentiation in hiPSCs differentiated on VN coated dishes in comparison to those differentiated on routinely used feeder cells. Analysis of PDX1 expression in differentiated cells illustrated that VN was able to further induce pancreatic progenitors as well. Altogether, our results demonstrated that VN efficiently promoted endodermal differentiation of human induced pluripotent stem cells.