Efficient derivation of functional astrocytes from human induced pluripotent stem cells (hiPSCs)

PLoS One. 2024 Dec 4;19(12):e0313514. doi: 10.1371/journal.pone.0313514. eCollection 2024.

Abstract

Astrocytes are specialized glial cell types of the central nervous system (CNS) with remarkably high abundance, morphological and functional diversity. Astrocytes maintain neural metabolic support, synapse regulation, blood-brain barrier integrity and immunological homeostasis through intricate interactions with other cells, including neurons, microglia, pericytes and lymphocytes. Due to their extensive intercellular crosstalks, astrocytes are also implicated in the pathogenesis of CNS disorders, such as ALS (amyotrophic lateral sclerosis), Parkinson's disease and Alzheimer's disease. Despite the critical importance of astrocytes in neurodegeneration and neuroinflammation are recognized, the lack of suitable in vitro systems limits their availability for modeling human brain pathologies. Here, we report the time-efficient, reproducible generation of astrocytes from human induced pluripotent stem cells (hiPSCs). Our hiPSC-derived astrocytes expressed characteristic astrocyte markers, such as GFAP, S100b, ALDH1L1 and AQP4. Furthermore, hiPSC-derived astrocytes displayed spontaneous calcium transients and responded to inflammatory stimuli by the secretion of type A1 and type A2 astrocyte-related cytokines.

MeSH terms

  • Aquaporin 4 / metabolism
  • Astrocytes* / cytology
  • Astrocytes* / metabolism
  • Cell Differentiation*
  • Cells, Cultured
  • Cytokines / metabolism
  • Glial Fibrillary Acidic Protein / metabolism
  • Humans
  • Induced Pluripotent Stem Cells* / cytology
  • Induced Pluripotent Stem Cells* / metabolism
  • S100 Calcium Binding Protein beta Subunit* / metabolism

Substances

  • S100 Calcium Binding Protein beta Subunit
  • Cytokines
  • Aquaporin 4
  • S100B protein, human
  • Glial Fibrillary Acidic Protein

Grants and funding

This study was supported by the Ministry of Health of the Czech Republic in the form of a grant [NU20-09-00437] to JL and RK, the National Research, Development and Innovation Fund in the form of a grant [2020-1.1.5-GYORSÍTÓSÁV-2021-00016] to AD, and the ERA-NET NEURON Research Programme in the form of a grant [2019-2.1.7-ERA-NET-2022-00031, NDCil project] to AD. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.