Abstract
Current advances in stem cell biology have brought much hope for therapy of neuro-degenerative diseases. However, neural stem cells (NSCs) are rare adult stem cells, and the use of non-NSCs requires efficient and high-yielding lineage-specific differentiation prior to transplantation for efficacy. We report on the efficient differentiation of human embryonic stem cell-derived mesenchymal progenitors (EMPs) into a neural phenotype with use of Y-27632, a clinically compliant small molecular inhibitor of Rho kinase (ROCKs). EMPs are a population of mesenchymal progenitors derived from human embryonic stem cells (hESCs) which are capable of multilineage differentiation but do not form teratomas. These early mesenchymal progenitors are an ideal stem cell source since they are non-tumorigenic and immunomodulatory—similar to adult bone marrow mesenchymal stem cells—but do not easily senesce after in vitro culture, and are able to be regenerated indefinitely along from ESCs and other pluripotent stem cells including induced pluripotent stem cells. We treated EMPs with Y-27632 and investigated for differentiation capacity and neural lineages by gene expression, and protein expression. Y-27632 induced EMPs into a neural-like morphology, with rapid development of cell extensions and processes within 24 hours. Y-27632-treated EMPs express several neural progenitor marker included Musashi1and neurogenin1 at the RNA level and GFAP at RNA and protein level. EMPs can differentiate into a neural phenotype via inhibition of RhoA/ROCKs pathway by Y-27632. Moreover, both in specific knockdown Rho/ROCKs pathway or inhibited downstream of ROCK II have the same effects of neural-like morphology. Taken together, our data indicate that inhibition of RhoA-kinase induce of neural phenotype of embryonic stem cell-derived mesenchymal progenitors.