Improving and accelerating the differentiation and functional maturation of human stem cell‐derived neurons: role of extracellular calcium and GABA. (15th November 2016)

• Record Type: Journal Article
• Authors: Kemp, Paul J.; Rushton, David J.; Yarova, Polina L.; Schnell, Christian; Geater, Charlene; Hancock, Jane M.; Wieland, Annalena; Hughes, Alis; Badder, Luned; Cope, Emma; Riccardi, Daniela; Randall, Andrew D.; Brown, Jonathan T.; Allen, Nicholas D.; Telezhkin, Vsevolod
• Is Part Of: Journal of physiology. Volume 594:Number 22(2016:Nov.)
• Page Start: 6583
• Page End: 6594
• Publication Date: 2016-11-15
• Subjects: Physiology -- Periodicals; 612.005
• DOI: 10.1113/JP270655
• Languages: English
• ISSNs: 0022-3751

Abstract

Abstract : Rationale and basic protocol for accelerating neuronal maturation using SynaptoJuice. Neural precursor cells (NPCs) that have been generated from either human embryonic stem (ES) cells or human induced pluripotent stem (iPS) cells are plated at day 0 in SynaptoJuice A. This first medium has been designed to synchronize neurogenesis, by forcing NPCs to exit the cell cycle, whilst increasing GABA‐induced Ca 2+ influx. At day 7, and thereafter, SynaptoJuice A is replaced by SynaptoJuice B, which is designed to promote synaptogenesis via WNT and TrkB signalling. At Day 21, the resulting neurons demonstrate punctate co‐registration of synaptic markers (synaptophysin and PSD95 are shown) and synchronized electrical activity (an exemplar spontaneous train of action potentials is shown). 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.