Generation of Human iPSC-Derived Astrocytes with a mature star-shaped phenotype for CNS modeling

• DOI: 10.1007/s12015-022-10376-2
• Publication Date: 2022-04-30T05:22:23Z
• Authors: Dimitrios Voulgaris; Polyxeni Nikolako...; Anna Herland
• Keywords: Neuroepithelial cell

Abstract The generation of astrocytes from human induced pluripotent stem cells has been hampered by either prolonged differentiation—spanning over two months—or shorter protocols that generate immature astrocytes, devoid salient mature astrocytic traits pivotal for central nervous system (CNS) modeling. We directed stable hiPSC-derived neuroepithelial to iPSC-derived Astrocytes (hiAstrocytes) with a high percentage star-shaped orchestrating an astrocytic-tuned culturing environment in 28 days. employed RT-qPCR and ICC validate the commitment cells. To evaluate inflammatory phenotype, we challenged hiAstrocytes pro-inflammatory cytokine IL-1β (interleukin 1 beta) quantitatively assessed secretion profile astrocyte-associated cytokines expression intercellular adhesion molecule (ICAM-1). Finally, capacity synthesize export antioxidant glutathione. In under days, generated express canonical markers, denoted GFAP, AQP4 ALDH1L1. addition, notion phenotype is reinforced both glutamate transporters EAAT1 EAAT2. Thus, have encompasses critical CNS modeling, including glutathione synthesis secretion, upregulation ICAM-1 on par fetal astrocytes. This protocol generates multifaceted model suitable vitro disease modeling personalized medicine. Graphical abstract