ABSTRACT
BACKGROUND: The in vitro differentiation methods of stem cell-derived dopaminergic neurons that serve as a cell source for the replacement therapy of Parkinson's disease are continuously optimized and improved, as well as the subsequent identification methods and testing indicators. OBJECTIVE: To observe the morphological development and electrophysiological characteristics of dopaminergic neurons differentiated from human embryonic stem cells so as to identify whether these differentiated cells have mature morphology and function under the current differentiation program. METHODS: Monolayer adherent method combined with dual-SMAD signaling inhibition was used to induce the directional differentiation of human embryonic stem cells into dopaminergic neurons. Then the cells were identified by light microscopy, electron microscopy and immunofluorescence, and the electrophysiological properties of dopaminergic neurons were detected by patch clamp electrophysiological technique. Herein, we evaluated the electrophysiological functions of dopaminergic neurons differentiated in vitro, with reference to the evaluation standard of dopaminergic neuron in vivo. RESULTS AND CONCLUSION: In this study, we successfully obtained dopaminergic neurons with mature morphology and functions differentiated from human embryonic stem cells in vitro. Findings from the subsequent electrophysiological test confirmed that dopaminergic neurons we acquired had electrophysiological properties in accordance with the evaluation standards of dopaminergic neurons in vivo. To conclude, the monolayer adherent method combined with dual-SMAD signaling inhibition can successfully induce the directional differentiation of human embryonic stem cells into dopaminergic neurons with mature morphology and functions.