| The proliferative capacity and differentiation potential of human embryonic stem cells (hESCs) makes them attractive for cell based therapies, including cell replacement therapy for Parkinson's disease. This study involved development of protocols essential for establishing a clinically acceptable hESC-derived dopaminergic neuron population. Undifferentiated hESCs were maintained in murine embryonic fibroblast-based and feeder-free culture for the first time in our facility. Expression of the pluripotency markers SSEA-4 and alkaline phosphatase suggested pluripotency. A purity of 39.1% and pluripotent cell fold expansion of 3.63x104 over 56 days was achieved. Differentiation of hESCs to dopaminergic neurons in static culture and differentiation to neural precursor cells (NPCs) in suspension culture through formation of embryoid bodies (EBs) and neurospheres followed. Formation of neurospheres produced the most promising result. Aggregates containing NPCs were present, determined by the presence of nestin-positive cells. Results of this study provide a solid foundation for future hESC-based research within our laboratory. |
