PMID- 25940691
OWN - NLM
STAT- MEDLINE
DCOM- 20160223
LR  - 20220310
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 5
DP  - 2015 May 5
TI  - Systematic optimization of human pluripotent stem cells media using Design of 
      Experiments.
PG  - 9834
LID - 10.1038/srep09834 [doi]
LID - 9834
AB  - 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 (NRG1beta1). 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.
FAU - Marinho, Paulo A
AU  - Marinho PA
AD  - 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.
FAU - Chailangkarn, Thanathom
AU  - Chailangkarn T
AD  - 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.
FAU - Muotri, Alysson R
AU  - Muotri AR
AD  - 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.
LA  - eng
GR  - DP2 OD006495/OD/NIH HHS/United States
GR  - 1-DP2-OD006495-01/OD/NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20150505
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Culture Media, Conditioned)
RN  - 0 (NRG1 protein, human)
RN  - 0 (Neuregulin-1)
RN  - 103107-01-3 (Fibroblast Growth Factor 2)
SB  - IM
MH  - Algorithms
MH  - Batch Cell Culture Techniques/*methods
MH  - Cell Differentiation/physiology
MH  - Cell Line
MH  - Cell Proliferation/physiology
MH  - Culture Media, Conditioned/chemistry/metabolism
MH  - Fibroblast Growth Factor 2/chemistry/*metabolism
MH  - Humans
MH  - Models, Statistical
MH  - Neuregulin-1/chemistry/*metabolism
MH  - Pluripotent Stem Cells/*cytology/*physiology
MH  - *Research Design
PMC - PMC4419516
EDAT- 2015/05/06 06:00
MHDA- 2016/02/26 06:00
PMCR- 2015/05/05
CRDT- 2015/05/06 06:00
PHST- 2014/10/23 00:00 [received]
PHST- 2015/03/17 00:00 [accepted]
PHST- 2015/05/06 06:00 [entrez]
PHST- 2015/05/06 06:00 [pubmed]
PHST- 2016/02/26 06:00 [medline]
PHST- 2015/05/05 00:00 [pmc-release]
AID - srep09834 [pii]
AID - 10.1038/srep09834 [doi]
PST - epublish
SO  - Sci Rep. 2015 May 5;5:9834. doi: 10.1038/srep09834.