Derivation, expansion and differentiation of induced pluripotent stem cells in continuous suspension cultures
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We describe derivation of induced pluripotent stem cells (iPSCs) from terminally differentiated mouse cells in serum- and feeder-free stirred suspension cultures. Temporal analysis of global gene expression revealed high correlations between cells reprogrammed in suspension and cells reprogrammed in adhesion-dependent conditions. Suspension culture-reprogrammed iPSCs (SiPSCs) could be differentiated into all three germ layers in vitro and contributed to chimeric embryos in vivo. SiPSC generation allowed for efficient selection of reprogramming factor-expressing cells based on their differential survival and proliferation in suspension culture. Seamless integration of SiPSC reprogramming and directed differentiation enabled scalable production of beating cardiac cells in a continuous single cell- and small aggregate-based process. This method is an important step toward the development of robust PSC generation, expansion and differentiation technology.
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A microfluidic neural tube device presented simultaneous opposing gradients of growth factors to direct neural tube patterning, enabling recapitulation of the in vivo structure.
“a microfluidic neural tube device was developed to present simultaneous opposing gradients of growth factors to direct neural tube patterning, enabling recapitulation of the in vivo structure92.”
From Organoids (2022) · cites this paper as [92]
The Introduction and Results describe continuous suspension culture and cardiac differentiation, while Methods > Differentiation protocols describes ectoderm differentiation without a neural tube device. Searches for microfluidic devices, opposing growth-factor gradients, and neural tube patterning found no passage addressing the attributed claim. The indexed passages provide no page numbers. (LLM, conf=0.76)
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- Authors
- Fluri, D. A., Tonge, P. D., Song, H., Baptista, R. P., Shakiba, N., Shukla, S., Clarke, G., Nagy, A., & Zandstra, P. W.
- Affiliations
- University of Toronto (CA)
- Mount Sinai Hospital (CA)
- University Health Network (CA)
- Corresponding author
- Zandstra, Peter W
- Venue
- Nature Methods
- Published
- 2012-03-25
- Volume
- 9
- Issue
- 5
- Pages
- 509-516
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- PMC
- PMC4954777
- OpenAlex
- W2015831230
- Cited by
- 106
- References
- 30
- Citations by year
- 2026: 2 · 2025: 1 · 2024: 6 · 2023: 3 · 2022: 4 · 2021: 3 · 2020: 3 · 2019: 3 · 2018: 8 · 2017: 6 · 2016: 12 · 2015: 12 · 2014: 17 · 2013: 17 · 2012: 9
- FWCI
- 7.08
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Methods (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Canadian Institutes of Health Research (57885_1, MOP57885)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Keywords
- induced pluripotent stem cells, suspension culture, iPSC derivation, iPSC differentiation, cardiac differentiation, beating cardiomyocytes, serum-free culture, feeder-free culture, reprogramming, chimeric embryos, three germ layers
- MeSH terms
- Animals, Cell Differentiation, Chimera, Fibroblasts, Cell Culture Techniques, Myocytes, Cardiac, Mice, Induced Pluripotent Stem Cells, Cellular Reprogramming
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