Improving and accelerating the differentiation and functional maturation of human stem cell‐derived neurons: role of ...

[Amanita Research]

Title: Improving and accelerating the differentiation and functional maturation of human stem cell‐derived neurons: role of extracellular calcium and GABA

Abstract: Neurons differentiated from pluripotent stem cells using established neural culture conditions often exhibit functional deficits. Recently, we have developed enhanced media which both synchronize the neurogenesis of pluripotent stem cell‐derived neural progenitors and accelerate their functional maturation; together these media are termed SynaptoJuice. This pair of media are pro‐synaptogenic and generate authentic, mature synaptic networks of connected forebrain neurons from a variety of induced pluripotent and embryonic stem cell lines. Such enhanced rate and extent of synchronized maturation of pluripotent stem cell‐derived neural progenitor cells generates neurons which are characterized by a relatively hyperpolarized resting membrane potential, higher spontaneous and induced action potential activity, enhanced synaptic activity, more complete development of a mature inhibitory GABAA receptor phenotype and faster production of electrical network activity when compared to standard differentiation media. This entire process – from pre‐patterned neural progenitor to active neuron – takes 3 weeks or less, making it an ideal platform for drug discovery and disease modelling in the fields of human neurodegenerative and neuropsychiatric disorders, such as Huntington's disease, Parkinson's disease, Alzheimer's disease and Schizophrenia.

Point of interest:
"...resulted in complete abolishment of the ACM‐evoked enhancements of spontaneous activity and significantly depolarized resting membrane potential (Fig. 1B). Furthermore, similar diminutions were seen when ionotropic GABAA receptors were inhibited using bicuculline, the first suggestion that a major contributor to Ca2+ influx during differentiation might be provided by excitatory GABAergic synaptic input in these developing neuronal networks".

"...by chronically raising the extracellular GABA concentration, and the [Ca2+]o to a level similar to that measured in fetal plasma, namely 1.8 mm (Kovacs & Kronenberg, 1997)".

https://physoc.onlinelibrary.wiley.com/ ... 3/JP270655