Cardiomyocyte Differentiation and Expansion

CF differentiation was induced in monolayer, similarly to Zhao et al. (2017) (link). Briefly, 25 × 10³ EPI (CTRL1, CTRL2, CTRL4 LQT1, ACM) were seeded per cm² on tissue culture plates coated with vitronectin in BPEL medium supplemented with FGF2 (10 ng/ml; R&D Systems) on day 12. On day 13 and every 2 days thereafter, medium was refreshed with BPEL supplemented with FGF2 (10 ng/ml). After 6 days (on day 19), CFs were expanded by changing BPEL to Fibroblast Growth Medium 3 (FGM3; PromoCell). FGM3 was refreshed every 2 days for approximately 10 days in total. After 10 days (on day 29), CFs were confluent and ready to be passaged at 1:2 ratio. FGM3 was refreshed the day after passaging and every 2 days thereafter. CFs (10cm² per vial) were cryopreserved in CryoStor CS10 medium (0.5ml/vial; Stem Cell Technologies).

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Giacomelli E., Meraviglia V., Campostrini G., Cochrane A., Cao X., van Helden R.W., Krotenberg Garcia A., Mircea M., Kostidis S., Davis R.P., van Meer B.J., Jost C.R., Koster A.J., Mei H., Míguez D.G., Mulder A.A., Ledesma-Terrón M., Pompilio G., Sala L., Salvatori D.C., Slieker R.C., Sommariva E., de Vries A.A., Giera M., Semrau S., Tertoolen L.G., Orlova V.V., Bellin M, & Mummery C.L. (2020). Human-iPSC-Derived Cardiac Stromal Cells Enhance Maturation in 3D Cardiac Microtissues and Reveal Non-cardiomyocyte Contributions to Heart Disease. Cell Stem Cell, 26(6), 862-879.e11.

Publication 2020

FGF2 Fibroblast Growth Medium 3 (FGM3; CryoStor CS10 medium

Fgf2 Fibroblast Lqt1 Stem cell Tissue Vitronectin

Corresponding Organization : University of Twente

Other organizations : Leiden University, Centro de Biología Molecular Severo Ochoa, Autonomous University of Madrid, Centro Cardiologico Monzino, University of Milan, Amsterdam UMC Location VUmc

Top 5 similar protocols

Variable analysis

independent variables

Induction of CF differentiation

dependent variables

Differentiation of EPI cells (CTRL1, CTRL2, CTRL4 LQT1, ACM) into CFs

control variables

Seeding density of 25 × 10^3 EPI cells per cm^2
Culture plates coated with vitronectin
BPEL medium supplemented with FGF2 (10 ng/ml) used for initial CF differentiation
Fibroblast Growth Medium 3 (FGM3) used for CF expansion

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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