Defined culture conditions robustly maintain human stem cell pluripotency, highlighting a role for Ca2+ signalingShow others and affiliations
2025 (English)In: Communications Biology, E-ISSN 2399-3642, Vol. 8, no 1, article id 255Article in journal (Refereed) Published
Abstract [en]
Induced pluripotent stem cells (iPSCs) have significant potential for disease modeling and cell therapies. However, their wide-spread application has faced challenges, including batch-to-batch variabilities, and notable distinctions when compared to embryonic stem cells (ESCs). Some of these disparities can stem from using undefined culture conditions and the reprogramming procedure, however, the precise mechanisms remain understudied. Here, we compared gene expression data from over 100 iPSC and ESC lines cultivated under undefined and defined conditions. Defined conditions significantly reduced inter-PSC line variability, irrespective of PSC cell type, highlighting the importance of standardization to minimize PSC biases. This variability is concurrent with decreased somatic cell marker and germ layer differentiation gene expression and increased Ca2+-binding protein expression. Moreover, SERCA pump inhibition highlighted an important role for intracellular Ca2+ activity in maintaining pluripotency gene expression under defined conditions. Further understanding of these processes can help standardize and improve defined hPSC culture conditions.
Place, publisher, year, edition, pages
Nature Portfolio , 2025. Vol. 8, no 1, article id 255
Keywords [en]
Induced pluripotent stem cells (iPSCs), Mutations
National Category
Cell and Molecular Biology
Research subject
Bioinformatics
Identifiers
URN: urn:nbn:se:his:diva-24929DOI: 10.1038/s42003-025-07658-zISI: 001424571300005PubMedID: 39966571Scopus ID: 2-s2.0-85219135139OAI: oai:DiVA.org:his-24929DiVA, id: diva2:1941261
Funder
Karolinska InstituteSwedish Research Council, 2019-01498The Swedish Brain Foundation, FO2019-0246The Swedish Brain Foundation, FO2021-0234Vinnova, IndiCell 2021-02695Swedish Cancer Society, 20 1159 PjSwedish Research Council, 2017-00815Swedish Research Council, 2021-03108The Swedish Brain Foundation, FO2018-0209The Swedish Brain Foundation, FO2020-0199Swedish Cancer Society, 19 0544 PjSwedish Cancer Society, 19 0545 UsSwedish Cancer Society, 22 2454 PjSwedish Childhood Cancer Foundation, PR2020-0124Swedish Childhood Cancer Foundation, PR2022-0111Vinnova, 2021-01834Swedish Society for Medical Research (SSMF), PG-22-0462
Note
CC BY 4.0
Correspondence and requests for materials should be addressed to Ilse Eidhof or Anna Falk.
Open access funding provided by Karolinska Institute.
We thank the donors, patients, and families, employees of the iPS core facility, Falk and Uhlén laboratory at Karolinska Institutet. We acknowledge WiCell, the Niklas Dahl laboratory at Uppsala University, and Fredrik Lanner laboratory at Karolinska Institute for providing PSC lines. We thank the BEA facility for Illumina array support. This study was financed by grants to A.F., VR (2019-01498), hjärnfonden (FO2019-0246, FO2021-0234), VINNOVA (IndiCell 2021-02695), cancerfonden (20 1159 Pj), and to P.U. (VR 2017-00815 and 2021-03108), Hjärnfonden (FO2018-0209 and FO2020-0199), Cancerfonden (19 0544 Pj, 19 0545 Us, and 22 2454 Pj), Barncancerfonden (PR2020-0124 and PR2022-0111). I.E. was supported by a MSCA EF Seal Of Excellence postdoctoral fellowship from VINNOVA (2021-01834) and a SSMF Postdoctoral Grant 2023 (PG-22-0462). We support inclusive, diverse, and equitable conduct of research.
2025-02-282025-02-282025-04-15Bibliographically approved