METHOD FOR DIFFERENTIATING PLURIPOTENT STEM CELLS INTO DOPAMINERGIC NERVE CELLS IN MIDBRAIN SUBSTANTIA NIGRA

摘要

Specific method for differentiating pluripotent stem cells into dopaminergic (A9 mDA) nerve cells in the midbrain substantia nigra is provided. Mature A9 mDA neurons are formed by differentiation, which can express the molecular markers of the midbrain substantia nigra dopaminergic neurons, including TH, FOXA2, EN1, LMX1A, NURR1 and GIRK2, but which rarely express the marker CALB of the ventral tegmental area dopaminergic neurons. A9 mDA nerve cells are transplanted into the substantia nigra, and the axons can project to the target brain area which is innervated by endogenous substantia nigra dopaminergic neurons, the dorsal striatum; the transplanted A9 mDA neurons themselves exhibit the classic electrophysiological characteristics of endogenous substantia nigra dopaminergic neurons, including a low-frequency spontaneous discharge frequency, and can induce sag by means of hyperpolarizing current stimulation, and transplanting the A9 mDA nerve cells into the substantia nigra or striatum of individuals with neurodegenerative diseases can alleviate motor deficits.