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PMID: 25940691 Published · epublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Systematic optimization of human pluripotent stem cells media using Design of Experiments.

Scientific reports ·Vol. 5 ·2015-05-05 ·Pages 9834

Marinho PA, Chailangkarn T, Muotri AR

Abstract

Human pluripotent stem cells (hPSC) are used to study the early stages of human development in vitro and, increasingly due to somatic cell reprogramming, cellular and molecular mechanisms of disease. Cell culture medium is a critical factor for hPSC to maintain pluripotency and self-renewal. Numerous defined culture media have been empirically developed but never systematically optimized for culturing hPSC. We applied design of experiments (DOE), a powerful statistical tool, to improve the medium formulation for hPSC. Using pluripotency and cell growth as read-outs, we determined the optimal concentration of both basic fibroblast growth factor (bFGF) and neuregulin-1 beta 1 (NRG1β1). The resulting formulation, named iDEAL, improved the maintenance and passage of hPSC in both normal and stressful conditions, and affected trimethylated histone 3 lysine 27 (H3K27me3) epigenetic status after genetic reprogramming. It also enhances efficient hPSC plating as single cells. Altogether, iDEAL potentially allows scalable and controllable hPSC culture routine in translational research. Our DOE strategy could also be applied to hPSC differentiation protocols, which often require numerous and complex cell culture media.

MeSH Terms
Algorithms Batch Cell Culture Techniques/methods Cell Differentiation/physiology Cell Line Cell Proliferation/physiology Culture Media, Conditioned/chemistry,metabolism Fibroblast Growth Factor 2/chemistry,metabolism Humans Models, Statistical Neuregulin-1/chemistry,metabolism Pluripotent Stem Cells/cytology,physiology Research Design
Chemicals
Culture Media, Conditioned NRG1 protein, human Neuregulin-1 Fibroblast Growth Factor 2
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Marinho Paulo A
University of California San Diego, School of Medicine, Department of Pediatrics/Rady Children's Hospital San Diego, Department of Cellular &Molecular Medicine, Stem Cell Program, La Jolla, CA 92093, MC 0695, USA.
Chailangkarn Thanathom
University of California San Diego, School of Medicine, Department of Pediatrics/Rady Children's Hospital San Diego, Department of Cellular &Molecular Medicine, Stem Cell Program, La Jolla, CA 92093, MC 0695, USA.
Muotri Alysson R
University of California San Diego, School of Medicine, Department of Pediatrics/Rady Children's Hospital San Diego, Department of Cellular &Molecular Medicine, Stem Cell Program, La Jolla, CA 92093, MC 0695, USA.
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Article Info
Journal
Scientific reports
Abbr.
Sci Rep
ISSN
2045-2322
DOI
10.1038/srep09834
Published
2015-05-05
Epub
2015-00-05
Pages
9834
Language
English
Region
England
NLM ID
101563288
PMCID
PMC4419516
Subset
IM
Grants
NIH HHS · DP2 OD006495 · United States
NIH HHS · 1-DP2-OD006495-01 · United States
External Links
PubMed source DOI: 10.1038/srep09834 PMC Full Text View journal details
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