the total live cell count obtained on day 12, good CM yields fall
around 1.9 106/mL [3, 9]. Percentage of cells beating falls
around 70% [12].
18. Attempt to break MC clumps and dissociate cells by pipetting
after 5 min incubation is complete. If MC clumps remain, place
back into 37 C for longer incubation. It can take up to 2 h for
cardiomyocytes to properly detach and dissociate from MC
clumps depending on the original hiPSC line used. If too
much pipetting is required to remove the cells or if clumps
remain stubborn, incubate it for more time at 37 C on a
shaker at 75 rpm.
References
1. Jafarkhani M, Salehi Z et al (2018) Strategies
for directing cells into building functional
hearts
and
parts.
Biomater
Sci
6
(7):1664–1690.
https://doi.org/10.1039/
c7bm01176h
2. Garbern JC, Lee RT (2013) Cardiac stem cell
therapy and the promise of heart regeneration.
Cell Stem Cell 12(6):689–698. https://doi.
org/10.1016/j.stem.2013.05.008
3. Laco F, Lam ATL et al (2020) Selection of
human induced pluripotent stem cells lines
optimization of cardiomyocytes differentiation
in an integrated suspension microcarrier biore-
actor. Stem Cell Res Ther 11:188. https://doi.
org/10.1186/s13287-020-01618-6
4. Ban K, Bae S et al (2017) Current strategies
and challenges for purification of cardiomyo-
cytes derived from human pluripotent stem
cells. Theranostics 7(7):2067–2077. https://
doi.org/10.7150/thno.19427
5. Chen VC, Ye J et al (2015) Development of a
scalable suspension culture for cardiac differen-
tiation from human pluripotent stem cells.
Stem Cell Res 15(2):365–375. https://doi.
org/10.1016/j.scr.2015.08.002
6. Serra M, Brito C et al (2012) Process engineer-
ing of human pluripotent stem cells for clinical
application.
Trends
Biotechnol
30
(6):350–359.
https://doi.org/10.1016/j.
tibtech.2012.03.003
7. Laco F, Woo TL et al (2018) Unraveling the
inconsistencies
of
cardiac
differentiation
efficiency induced by the GSK3beta inhibitor
CHIR99021 in human pluripotent stem cells.
Stem Cell Rep 10(6):1851–1866. https://doi.
org/10.1016/j.stemcr.2018.03.023
8. Ting S, Chen A et al (2014) An intermittent
rocking platform for integrated expansion and
differentiation of human pluripotent stem cells
to cardiomyocytes in suspended microcarrier
cultures.
Stem
Cell
Res
13(2):202–213.
https://doi.org/10.1016/j.scr.2014.06.00
9. Ting S, Lam A (2018) Meticulous optimiza-
tion of cardiomyocyte yields in a 3-stage con-
tinuous integrated agitation bioprocess. Stem
Cell Res 31:161–173. https://doi.org/10.
1016/j.scr.2018.07.020
10. Chen AK, Chen X et al (2011) Critical micro-
carrier properties affecting the expansion of
undifferentiated human embryonic stem cells.
Stem Cell Res 7(2):97–111. https://doi.org/
10.1016/j.scr.2011.04.007
11. Lam AT, Chen AK (2014) Conjoint propaga-
tion and differentiation of human embryonic
stem cells to cardiomyocytes in a defined
microcarrier spinner culture. Stem Cell Res
Ther
5(5):110.
https://doi.org/10.1186/
scrt498
12. Lecina M, Ting S et al (2010) Scalable platform
for human embryonic stem cell differentiation
to cardiomyocytes in suspended microcarrier
cultures.
Tissue
Eng
16(6):1609–1619.
https://doi.org/10.1089/ten.TEC.2010.
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