Directed differentiation of functional astroglial subtypes from human pluripotent stem cells

Abstract

Regionally and functionally diverse types of astrocytes exist throughout the central nervous system and participate in nearly every aspect of normal and abnormal neural function. Therefore, human astrocyte subtypes are a useful tool for understanding brain function, modulating disease processes, and promoting neural regeneration. Here we describe a protocol for directed differentiation and maintenance of functional astroglia from human pluripotent stem cells in a chemically defined system. Human stem cells are first differentiated to neuroepithelial cells with or without exogenous patterning molecules(days 0 –21). Regular dissociation of the neuroepithelial clusters in suspension, and in the presence of mitogens, permits generation of astroglial subtypes over along term expansion (days 21–90). Finally, the astroglial progenitors are either amplified for an extended time or differentiated to functional astrocytes upon removal of mitogens and the addition of CNTF ( days > 90). This method generates robust populations of functionally diversified astrocytes with high efficiency.

Human iPSC-Derived Neural Progenitors Are an Effective Drug Discovery Model for Neurological mtDNA Disorders

“…The differentiation of NI NPCs into astrocytes was promoted by a standardized protocol (Krencik and Zhang, 2011) with slight modifications. In brief, NPCs were cultured as neurospheres in uncoated flasks with NI medium for 3 months during which progenitor clusters switched from neurogenic neurospheres to gliogenic astrospheres.…”

Transplantation of stem cell-derived astrocytes for the treatment of amyotrophic lateral sclerosis and spinal cord injury

“…These cells can be used to model human pathological processes in vitro and in vivo as well as for therapeutic transplantation [163] . More recently the Zhang laboratory reported another shorter duration protocol for generating astrocytes from iPSCs by differentiation of these cells into neuroepithelial cells, followed by removal of mitogen and treatment with CNTF to induce differentiation into astrocytes [70] . Recently, the Maragakis lab conducted transplantation of iPSC-and ESC-derived immature astrocytes into rodent spinal cord.…”

“…Astrocytes generated by this protocol were characterized by specific gene expression profiles, neurotransmitter and ionic uptake, support of neuronal maturation, and importantly expression of specific sets of homeodomain transcription factors that specify functional identity [69] . On the basis of this, another protocol was recently reported for an efficient generation of immature astrocytes from human pluripotent stem cells (hPSCs) [70] . The protocol consists of three steps.…”

Astrocytic transporters in Alzheimer's disease

“…In recent years investigators have developed methods to derive and maintain astrocytes from patient iPSCs (261)(262)(263). The question as to whether these cells truly represent functioning astrocytes in aged human brains remains to be answered (264).…”