Human CSCs were derived as previously descried using the cardiosphere
method.44 (link) In brief, heart tissues were
cut into tiny pieces and washed with PBS and digestion of collagenase
(Sigma, St. Louis, MO). Tissue fragments were cultured as “cardiac
explants” on a plate coated with 0.5 mg/mL fibronectin (BD
Biosciences, San Jose, CA) in IMDM supplemented with FBS, 0.5% gentamicin
(Gibco, Life Technologies, California, USA), 0.1 mM 2-mercaptoethanol
(Invitrogen), and 1%l -glutamine (Invitrogen). After about
7–14 days, we collected cardiac explanted-derived cells with
0.25% trypsin (Gibco) and then seeded them in ultralow attachment
flasks (Corning) for cardiospheres. After several days, cardiosphere-derived
cardiac stem cells were formed by seeding harvested cardiospheres
on fibronectin-coated plates and being incubated in 5% CO2 at 37 °C.
method.44 (link) In brief, heart tissues were
cut into tiny pieces and washed with PBS and digestion of collagenase
(Sigma, St. Louis, MO). Tissue fragments were cultured as “cardiac
explants” on a plate coated with 0.5 mg/mL fibronectin (BD
Biosciences, San Jose, CA) in IMDM supplemented with FBS, 0.5% gentamicin
(Gibco, Life Technologies, California, USA), 0.1 mM 2-mercaptoethanol
(Invitrogen), and 1%
7–14 days, we collected cardiac explanted-derived cells with
0.25% trypsin (Gibco) and then seeded them in ultralow attachment
flasks (Corning) for cardiospheres. After several days, cardiosphere-derived
cardiac stem cells were formed by seeding harvested cardiospheres
on fibronectin-coated plates and being incubated in 5% CO2 at 37 °C.
