Premium
GFAP‐independent inflammatory competence and trophic functions of astrocytes generated from murine embryonic stem cells
Author(s) -
Kuegler Philipp B.,
Baumann Bettina A.,
Zimmer Bastian,
Keller Sascha,
Marx Andreas,
Kadereit Suzanne,
Leist Marcel
Publication year - 2012
Publication title -
glia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.954
H-Index - 164
eISSN - 1098-1136
pISSN - 0894-1491
DOI - 10.1002/glia.21257
Subject(s) - biology , astrocyte , microglia , microbiology and biotechnology , glial fibrillary acidic protein , embryonic stem cell , induced pluripotent stem cell , stem cell , neural stem cell , neuroglia , immunology , neuroscience , inflammation , central nervous system , biochemistry , immunohistochemistry , gene
The directed generation of pure astrocyte cultures from pluripotent stem cells has proven difficult. Generation of defined pluripotent‐stem‐cell derived astrocytes would allow new approaches to the investigation of plasticity and heterogeneity of astrocytes. We here describe a two‐step differentiation scheme resulting in the generation of murine embryonic stem cell (mESC) derived astrocytes (MEDA), as characterized by the upregulation of 19 astrocyte‐associated mRNAs, and positive staining of most cells for GFAP (glial fibrillary acidic protein), aquaporin‐4 or glutamine synthetase. The MEDA cultures could be cryopreserved, and they neither contained neuronal, nor microglial cells. They also did not react to the microglial stimulus lipopolysaccharide, while inflammatory activation by a complete cytokine mix (CCM) or its individual components (TNF‐α, IL1‐β, IFN‐γ) was readily observed. MEDA, stimulated by CCM, became susceptible to CD95 ligand‐induced apoptosis and produced NO and IL‐6. This was preceded by NF‐kB activation, and up‐regulation of relevant mRNAs. Also GFAP‐negative astrocytes were fully inflammation‐competent. Neurotrophic support by MEDA was found to be independent of GFAP expression. In summary, we described here the generation and functional characterization of microglia‐free murine astrocytes, displaying phenotypic heterogeneity as is commonly observed in brain astrocytes. © 2011 WiWiley Periodicals, Inc.