TY - JOUR
T1 - Comparison of two protocols for the generation of iPSC-derived human astrocytes
AU - Mulica, Patrycja
AU - Venegas, Carmen
AU - Landoulsi, Zied
AU - Badanjak, Katja
AU - Delcambre, Sylvie
AU - Tziortziou, Maria
AU - Hezzaz, Soraya
AU - Ghelfi, Jenny
AU - Smajic, Semra
AU - Schwamborn, Jens
AU - Krüger, Rejko
AU - Antony, Paul
AU - May, Patrick
AU - Glaab, Enrico
AU - Grünewald, Anne
AU - Pereira, Sandro L
N1 - Funding
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 ATTRAC T 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
© 2023. BioMed Central Ltd., part of Springer Nature.
PY - 2023/9/20
Y1 - 2023/9/20
N2 - 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.
AB - 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.
UR - https://pubmed.ncbi.nlm.nih.gov/37730545
U2 - 10.1186/s12575-023-00218-x
DO - 10.1186/s12575-023-00218-x
M3 - Article
C2 - 37730545
SN - 1480-9222
VL - 25
JO - Biological Procedures Online
JF - Biological Procedures Online
IS - 1
M1 - 26
ER -