PMID- 32155214
OWN - NLM
STAT- MEDLINE
DCOM- 20200619
LR  - 20210402
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 15
IP  - 3
DP  - 2020
TI  - Practical adoption of state-of-the-art hiPSC-cardiomyocyte differentiation 
      techniques.
PG  - e0230001
LID - 10.1371/journal.pone.0230001 [doi]
LID - e0230001
AB  - Human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes are a valuable 
      resource for cardiac therapeutic development; however, generation of these cells 
      in large numbers and high purity is a limitation in widespread adoption. Here, 
      design of experiments (DOE) is used to investigate the cardiac differentiation 
      space of three hiPSC lines when varying CHIR99027 concentration and cell seeding 
      density, and a novel image analysis is developed to evaluate plate coverage when 
      initiating differentiation. Metabolic selection via lactate purifies 
      hiPSC-cardiomyocyte populations, and the bioenergetic phenotype and engineered 
      tissue mechanics of purified and unpurified hiPSC-cardiomyocytes are compared. 
      Findings demonstrate that when initiating differentiation one day after hiPSC 
      plating, low (3 muM) Chiron and 72 x 103 cells/cm2 seeding density result in peak 
      cardiac purity (50-90%) for all three hiPSC lines. Our results confirm that 
      metabolic selection with lactate shifts hiPSC-cardiomyocyte metabolism towards 
      oxidative phosphorylation, but this more "mature" metabolic phenotype does not by 
      itself result in a more mature contractile phenotype in engineered cardiac 
      tissues at one week of culture in 3D tissues. This study provides widely 
      adaptable methods including novel image analysis code and parameters for refining 
      hiPSC-cardiomyocyte differentiation and describes the practical implications of 
      metabolic selection of cardiomyocytes for downstream tissue engineering 
      applications.
FAU - Rupert, Cassady E
AU  - Rupert CE
AD  - Center for Biomedical Engineering, School of Engineering and Division of Biology 
      and Medicine, Brown University, Providence, RI, United States of America.
FAU - Irofuala, Chinedu
AU  - Irofuala C
AD  - Center for Biomedical Engineering, School of Engineering and Division of Biology 
      and Medicine, Brown University, Providence, RI, United States of America.
FAU - Coulombe, Kareen L K
AU  - Coulombe KLK
AUID- ORCID: 0000-0001-6664-339X
AD  - Center for Biomedical Engineering, School of Engineering and Division of Biology 
      and Medicine, Brown University, Providence, RI, United States of America.
LA  - eng
GR  - R01 HL135091/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20200310
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
SB  - IM
MH  - Adult
MH  - *Cell Differentiation
MH  - Cell Line
MH  - Cytological Techniques/*methods
MH  - Humans
MH  - Induced Pluripotent Stem Cells/*cytology
MH  - Myocytes, Cardiac/*cytology/metabolism
MH  - Phenotype
PMC - PMC7064240
COIS- The authors have declared that no competing interests exist.
EDAT- 2020/03/11 06:00
MHDA- 2020/06/20 06:00
PMCR- 2020/03/10
CRDT- 2020/03/11 06:00
PHST- 2019/11/26 00:00 [received]
PHST- 2020/02/19 00:00 [accepted]
PHST- 2020/03/11 06:00 [entrez]
PHST- 2020/03/11 06:00 [pubmed]
PHST- 2020/06/20 06:00 [medline]
PHST- 2020/03/10 00:00 [pmc-release]
AID - PONE-D-19-32871 [pii]
AID - 10.1371/journal.pone.0230001 [doi]
PST - epublish
SO  - PLoS One. 2020 Mar 10;15(3):e0230001. doi: 10.1371/journal.pone.0230001. 
      eCollection 2020.