GFAP-independent inflammatory competence and trophic functions of astrocytes generated from murine embryonic stem cells
| Autor: | Marcel Leist, Andreas Marx, Suzanne Kadereit, Bastian Zimmer, Sascha Keller, Philipp B. Kuegler, Bettina A. Baumann |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Primary Cell Culture
Cell Culture Techniques Cell Line Mice 03 medical and health sciences Cellular and Molecular Neuroscience chemistry.chemical_compound 0302 clinical medicine ddc:570 Glutamine synthetase Glial Fibrillary Acidic Protein medicine Animals Cell Lineage Nerve Growth Factors Induced pluripotent stem cell Embryonic Stem Cells 030304 developmental biology Inflammation Mice Inbred BALB C IL-6 0303 health sciences biology Glial fibrillary acidic protein glutamine synthetase Cell Differentiation Embryonic stem cell single cell Cell biology aquaporin Phenotype medicine.anatomical_structure Neurology Biochemistry chemistry Cell culture Astrocytes neuronal support biology.protein 030217 neurology & neurosurgery Neurotrophin MEDA Astrocyte |
| Zdroj: | Glia Glia; Vol 60 |
| ISSN: | 0894-1491 |
| DOI: | 10.1002/glia.21257 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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