Neuroinflammatory astrocytes generated from cord blood-derived human induced pluripotent stem cells.

ABSTRACT: BACKGROUND:Astrocytes respond to central nervous system (CNS) injury and disease by transforming to a reactive astrogliosis cell state that can contribute to either CNS dysfunction or repair. Neuroinflammation is a powerful driver of a harmful A1 astrogliosis phenotype associated with in vitro neurotoxicity and histopathology in human neurodegenerative diseases. Here we report a protocol for the rapid development of a human cell culture model of neuroinflammatory astrogliosis using induced pluripotent stem cells (iPSCs). METHODS:Using RNA sequencing and in vitro cell assays, we measured transcriptional and cellular effects of chronic exposure of human iPSC-derived astrocytes to the cytokines TNF? (tumor necrosis factor alpha) or IL-1? (interleukin-1 beta). RESULTS:We show TNF? and IL-1? induce pro-inflammatory gene signatures but by widely different magnitudes. TNF? treatment results in 606 differential expressed genes, the suppression of glutamate-uptake, and increased phagocytic activity in astrocyte cultures. In contrast, IL-1? effects are attenuated to 33 differential expressed genes and no significant effects on glutamate-uptake or increased phagocytic activity. CONCLUSION:Our approach demonstrates a rapid tool for modeling neuroinflammatory human astrocytic responses in nervous system trauma and disease. In particular, we reveal a model for robust TNF?-induced human astrogliosis suitable for the study of neurotoxic A1 astrocytes.

SUBMITTER: Zhou Q 

PROVIDER: S-EPMC6688278 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature