Rapid and efficient induction of functional astrocytes from human pluripotent stem cells

Abstract

The derivation of astrocytes from human pluripotent stem cells is currently slow and inefficient. We demonstrate that overexpression of the transcription factors SOX9 and NFIB in human pluripotent stem cells rapidly and efficiently yields homogeneous populations of induced astrocytes. In our study these cells exhibited molecular and functional properties resembling those of adult human astrocytes and were deemed suitable for disease modeling. Our method provides new possibilities for the study of human astrocytes in health and disease.

Plasminogen binding inhibitors demonstrate unwanted activities on GABA A and glycine receptors in human iPSC derived neurons

“…Pro-inflammatory response is used to validate model functionality of hPSC derived astrocytes. Increased IL6 and IL8 secretion in response to IL1B stimulation is observed for several hPSC derived astrocytic models (183,(242)(243)(244)(245) as well as to TNF (242,244,245) and Ab42 (246) stimulation. This is in concordance with pro-inflammatory stimulation of human primary fetal astrocytes (244,246).…”

“…Evaluation of calcium signaling is performed to functionally validate and assess the astrocyte biology of hPSC derived astrocytic models. Functional calcium signaling is shown in several hPSC derived models in response to mechanical (15,16,230,242,244), electrical (245), ATP (20,183,230) and glutamate simulation (246). Spontaneous calcium waves are also observed in hPSC derived models (183) together with calcium leakage from the ER, identified by inhibiting the major ER release channels RYR, IP3R and SERCA (245).…”

“…Astrocyte functionality is diverse and several astrocyte-associated functions besides glutamate uptake, inflammatory response and calcium signaling are studied using hPSC derived models including; potassium conductance (15,261), perineural net formation (262), oxygen consumption rates (245), phagocytosis (246), gap junctions connectivity (183) and metabolism (234). Notably, the development of astrocyte buffering functions of different ions is likely to be affected by medium composition where an imbalance might hinder this process.…”

“…This can be observed in neuronal cultures where classical basic medium and FBS impair electrical activity and synaptic communication (263). Moreover, hPSC derived astrocytes are being used to study astrocytic effects on neuronal biology including; synaptogenesis (183), neurite outgrowth (16,20) and neuronal electrophysiology (183). Additionally, assessment of human isolated and PSC derived astroglia biology is studied by engraftment in rodents (8,16,29,30,183).…”

An update on human astrocytes and their role in development and disease

“…The most conventional approach, 3D generation and maintenance, has become the standard method on which the majority of subsequent protocols have been based, including significant portions of direct conversion methods (Juopperi et al, 2012;Krencik & Zhang, 2011;Mormone, D'Sousa, Alexeeva, Bederson, & Germano, 2014). Finally, direct conversion through TF expression significantly decreases the induction protocols length but stands as a relatively new and unexplored approach (Caiazzo et al, 2014;Canals et al, 2018;Li, Tao, et al, 2018;Li, Tian, et al, 2018;Tchieu et al, 2019).…”