The hiPSCs were differentiated into cardiomyocytes as previously described (Campostrini et al., 2021 (link)). One day prior to differentiation (d-1), the hiPSCs were harvested using TrypLE Select and plated onto Matrigel (1:100, Corning)-coated wells in StemFlex™ Medium containing RevitaCell™ Supplement (1:200 dilution). On d0, the cells were refreshed with mBEL medium containing 5 μM CHIR99021 (Axon Medchem). On differentiation d2, the cells were refreshed with mBEL medium containing 5 μM of XAV939 (Tocris) and 0.25 μM IWP-L6 (AbMole). From differentiation d4 on, the cells were maintained in mBEL medium. The hiPSC-CMs were cryopreserved at differentiation d20 or d21 as previously described in a freezing medium comprising of 90% Knockout Serum Replacement (Gibco) and 10% DMSO (Brink et al., 2020 (link)). Subsequent thawing and seeding of the cells were performed as previously described (Brink et al., 2020 (link); Campostrini et al., 2021 (link)).
Iwp l6
Manufactured by AbMole
The IWP-L6 is a laboratory instrument designed for weighing purposes. It features a high-precision balance mechanism and a digital display for accurate measurements.
Automatically generated - may contain errors
Lab products found in correlation
Chir99021, by Axon Medchem (3 mentions)
Essential 8 medium, by Thermo Fisher Scientific (2 mentions)
Tryple select, by Thermo Fisher Scientific (2 mentions)
Stemflex medium, by Thermo Fisher Scientific (2 mentions)
Essential 8, by Thermo Fisher Scientific (2 mentions)
Revitacell supplement, by Thermo Fisher Scientific (2 mentions)
Xav939, by AbMole (2 mentions)
Matrigel, by Corning (1 mentions)
Knockout serum replacement (ksr), by Thermo Fisher Scientific (1 mentions)
Xav939, by Bio-Techne (1 mentions)
3 protocols using iwp l6
1
Differentiation and Cryopreservation of hiPSC-CMs
2
Cardiomyocyte Differentiation of hiPSCs
3
Cardiomyocyte Differentiation of hiPSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Subclones from two control hiPSC lines (LUMC0020iCTRL (Zhang et al., 2014 (link)) [LUMC20]; LUMC0099iCTRL [LUMC99]), and two hiPSC lines derived from patients diagnosed with HCM (LUMC0033iMyBPC [HCM1]; LUMC0035iMyBPC [HCM3] (Birket et al., 2015 (link))) were maintained either in Essential 8 or StemFlex Medium (both Gibco). One day prior to differentiation (d-1), the hiPSCs were harvested using TrypLE Select (Gibco) and plated onto Matrigel-coated wells of a 12-well cell culture plate, either in Essential 8 Medium containing RevitaCell Supplement (1:200 dilution; Gibco) or StemFlex Medium at 3.9 × 104/cm2. The hiPSCs were differentiated into cardiomyocytes either using the Pluricyte Cardiomyocyte Differentiation Kit (NCardia) according to the manufacturer's instructions, or in a modified BPEL medium (Elliott et al., 2011 (link)) supplemented with small molecules. Specifically, 5 μM CHIR99021 (Axon Medchem) from day 0 to day 2 of differentiation and 5 μM XAV939 + 0.25 μM IWP-L6 (AbMole) from differentiation day 2 to day 4. The hiPSC-CMs were maintained in Medium C (NCardia) until differentiation day 20–21 (LUMC20 and LUMC99), or the modified BPEL medium until differentiation day 14 or 17 (HCM1 and HCM3), and then dissociated as previously described (van den Berg et al., 2014 ).
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