Human iPSC cell line ACS-1021 (ATCC, USA) and CPCs induced by ISX-9 were cultured as described [15 (link)]. In some cases, EBs and commercial human CPCs were also cultured. Conditioned media was collected and EVs were isolated by centrifugation at 3000 rpm for 30 min to remove cells and debris, followed by filtration through a 0.22 μm filter to remove the remaining debris. Then, the medium was further concentrated to 500 μl using Amicon Ultra-15 100 kDa centrifugal filter units (Millipore). Isolation of EV in the concentrated medium was carried out through qEV size exclusion columns (Izon Science). EV fractions were collected and concentrated by Amicon Ultra-4 10 kDa centrifugal filter units to a final volume of <100 μl. The purified EVs were stored at -80°C and subsequently characterized by particle size, EV markers, and electron microscopy.
Acs 1021
Manufactured by American Type Culture Collection
Sourced in United States
The ACS-1021 is a benchtop autoclave designed for sterilizing laboratory equipment and media. It features a stainless steel chamber, digital controls, and a programmable cycle. The core function of the ACS-1021 is to provide a reliable and consistent method for sterilizing materials in a laboratory setting.
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Lab products found in correlation
2 protocols using acs 1021
1
Isolation and Characterization of Extracellular Vesicles from Cardiac Progenitor Cells
2
Feeder-Free Cardiac Differentiation of hiPSCs
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An iPSC line derived from human cardiac fibroblasts was purchased from ATCC (ACS-1021). We optimized the feeder free monolayer differentiation protocol as previously described (Bhattacharya et al., 2014 (link)). Briefly, the hiPSCs were plated on Geltrex (A1413202, Thermo Fisher)-coated six-well tissue culture dishes and cultured in Essential 8 Flex medium. For cardiomyocyte differentiation, hiPSCs were plated at a density of 100,000 cells/well, coated with Geltrex, and cultured in Essential 8 medium supplemented with Rho Kinase inhibitor, Y-27632 (1254, R&D Systems). After 24 hr the Rho Kinase inhibitor was removed and the cells were cultured for 4 days (Figure 1 A). On day 0, the cultures were initiated for differentiation using RPMI with B27 minus insulin (CM differentiation media) containing 6 μM CHIR99021 (4423, R&D Systems) for 48 hr. On day 3, the cultures were treated with 5 μM IWR-1 (I0161, Sigma Aldrich) for 48 hr. Subsequently, the cultures were maintained in cardiomyocyte differentiation media until day 10. Spontaneously beating cultures were observed around day 7 and hiPSC-CMs were cryopreserved at day 10.
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