Comparison of two protocols for the generation of iPSC-derived human astrocytes

Patrycja Mulica, Carmen Venegas, Zied Landoulsi, Katja Badanjak, Sylvie Delcambre, Maria Tziortziou, Soraya Hezzaz, Jenny Ghelfi, Semra Smajic, Jens Schwamborn, Rejko Krüger, Paul Antony, Patrick May, Enrico Glaab, Anne Grünewald*, Sandro L Pereira

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review


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.

Original languageEnglish
Article number26
JournalBiological Procedures Online
Issue number1
Publication statusPublished - 20 Sept 2023


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