Astrocytes; Disease modeling; iPSC; Biochemistry, Genetics and Molecular Biology (all); General Biochemistry, Genetics and Molecular Biology
Abstract :
[en] [en] BACKGROUND: Astrocytes have recently gained attention as key contributors to the pathogenesis of neurodegenerative disorders including Parkinson's disease. To investigate human astrocytes in vitro, numerous differentiation protocols have been developed. However, the properties of the resulting glia are inconsistent, which complicates the selection of an appropriate method for a given research question. Thus, we compared two approaches for the generation of iPSC-derived astrocytes. We phenotyped glia that were obtained employing a widely used long, serum-free ("LSF") method against an in-house established short, serum-containing ("SSC") protocol which allows for the generation of astrocytes and midbrain neurons from the same precursor cells.
RESULTS: We employed high-content confocal imaging and RNA sequencing to characterize the cultures. The astrocytes generated with the LSF or SSC protocols differed considerably in their properties: while the former cells were more labor-intense in their generation (5 vs 2 months), they were also more mature. This notion was strengthened by data resulting from cell type deconvolution analysis that was applied to bulk transcriptomes from the cultures to assess their similarity with human postmortem astrocytes.
CONCLUSIONS: Overall, our analyses highlight the need to consider the advantages and disadvantages of a given differentiation protocol, when designing functional or drug discovery studies involving iPSC-derived astrocytes.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
MULICA, Patrycja ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine > Molecular and Functional Neurobiology > Team Anne GRÜNEWALD
Venegas, Carmen; Luxembourg Centre for Systems Biomedicine, University of Luxembourg, Esch-Sur-Alzette, Luxembourg
LANDOULSI, Zied ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > Bioinformatics Core
BADANJAK, Katja ; University of Luxembourg > Faculty of Science, Technology and Medicine (FSTM) > FSTM Faculty administration > Research Facilitators
DELCAMBRE, Sylvie ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine > Molecular and Functional Neurobiology > Team Anne GRÜNEWALD
TZIORTZIOU, Maria ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > Molecular and Functional Neurobiology
HEZZAZ, Soraya ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > Molecular and Functional Neurobiology
Ghelfi, Jenny; Luxembourg Centre for Systems Biomedicine, University of Luxembourg, Esch-Sur-Alzette, Luxembourg
SMAJIC, Semra ; University of Luxembourg > Faculty of Science, Technology and Medicine (FSTM) > Department of Life Sciences and Medicine (DLSM) > Medical Education
SCHWAMBORN, Jens Christian ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > Developmental and Cellular Biology
Krüger, Rejko; Luxembourg Centre for Systems Biomedicine, University of Luxembourg, Esch-Sur-Alzette, Luxembourg ; Luxembourg Institute of Health, Strassen, Luxembourg
ANTONY, Paul ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > Scientific Central Services > Imaging Platform
MAY, Patrick ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > Bioinformatics Core
GLAAB, Enrico ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > Biomedical Data Science
GRÜNEWALD, Anne ✱; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > Molecular and Functional Neurobiology ; University of Lübeck > Institute of Neurogenetics
Pereira, Sandro L ✱; Luxembourg Centre for Systems Biomedicine, University of Luxembourg, Esch-Sur-Alzette, Luxembourg
Luxembourg National Research Fund Michael J Fox Foundation European Union’s Horizon Europe research and innovation program European Union’s Horizon 2020 research and innovation program Luxembourg National Research Fund (FNR), NextImmune2
Funding text :
PM and SS were supported by the Luxembourg National Research Fund (FNR) within the framework of the PARK-QC DTU (PRIDE17/12244779/PARK-QC). KB was supported by the FNR within the framework of the CriTICS DTU (FNR10907093). CV was funded by an FNR CORE Junior Grant (“NeuroFlame”, C20/BM/14548100). AG received funding through an FNR ATTRACT grant (“Model-IPD”, FNR9631103). AG, ZL, PM, JG, and SH were supported by the FNR within the INTER grant “ProtectMove” (INTER/DFG/19/14429377). In addition, AG, SLP and SD were supported by the FNR within the framework of a CORE grant (“CAMeSyn”, C19/BM/13688526). MT was supported by the FNR within the framework of the NextImmune2 DTU (PRIDE21/16749720). The work of RK was supported by the FNR within the following projects: MotaSYN and MAMaSyn. Moreover, RK is supported by the Michael J. Fox Foundation and the European Union’s Horizon Europe research and innovation program (Orchestra). AG, JS, RK and EG received support from the FNR as part of the National Centre for Excellence in Research on Parkinson’s disease (NCER-PD). Finally, EG was supported by the European Union’s Horizon 2020 research and innovation program under the grant no. ERAPERMED 2020–314 for the project DIGI-PD.We would like to thank Dr. Marja Koskuvi and Prof. Jari Koistinaho for the practical introduction to the Oksanen protocol. In addition, we thank the group of Prof. Jens Schwamborn, Center for Regenerative Therapies at TU Dresden, Integrated Biobank of Luxembourg, Max Planck Institute for Molecular Biomedicine in Münster and Hertie Institute for Clinical Brain Research in Tübingen for providing the cell lines used in the study.
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