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Defined culture conditions robustly maintain human stem cell pluripotency, highlighting a role for Ca2+ signaling

Eidhof, Ilse LU ; Ulfenborg, Benjamin ; Kele, Malin ; Shahsavani, Mansoureh ; Winn, Dania LU ; Uhlén, Per and Falk, Anna LU orcid (2025) In Communications Biology 8(1).
Abstract

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... (More)

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.

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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Communications Biology
volume
8
issue
1
article number
255
publisher
Nature Publishing Group
external identifiers
  • pmid:39966571
  • scopus:85219135139
ISSN
2399-3642
DOI
10.1038/s42003-025-07658-z
language
English
LU publication?
yes
id
13c81b39-138e-4138-b220-ffa905511bc3
date added to LUP
2025-06-09 10:48:44
date last changed
2025-07-07 13:41:25
@article{13c81b39-138e-4138-b220-ffa905511bc3,
  abstract     = {{<p>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 Ca<sup>2+</sup>-binding protein expression. Moreover, SERCA pump inhibition highlighted an important role for intracellular Ca<sup>2+</sup> activity in maintaining pluripotency gene expression under defined conditions. Further understanding of these processes can help standardize and improve defined hPSC culture conditions.</p>}},
  author       = {{Eidhof, Ilse and Ulfenborg, Benjamin and Kele, Malin and Shahsavani, Mansoureh and Winn, Dania and Uhlén, Per and Falk, Anna}},
  issn         = {{2399-3642}},
  language     = {{eng}},
  number       = {{1}},
  publisher    = {{Nature Publishing Group}},
  series       = {{Communications Biology}},
  title        = {{Defined culture conditions robustly maintain human stem cell pluripotency, highlighting a role for Ca<sup>2+</sup> signaling}},
  url          = {{http://dx.doi.org/10.1038/s42003-025-07658-z}},
  doi          = {{10.1038/s42003-025-07658-z}},
  volume       = {{8}},
  year         = {{2025}},
}