Culturing Human Fibroblasts and Induced Pluripotent Stem Cells

Human fetal fibroblasts (IMR90) were cultured in Dulbecco’s modified Eagle’s medium-high glucose (Invitrogen, Carlsbad, CA), 2 mM L-glutamine (Invitrogen), and 10% fetal bovine serum (Invitrogen). Cells were maintained at 37°C and 5% CO₂. Cells were passaged with 0.05% trypsin-ethylene diamine tetraacetic acid (Invitrogen). Skin fibroblasts from an FH patient (GM03040; Coriell Cell Repositories) were cultured in FH growth medium comprised of minimum essential medium (Invitrogen), 15% fetal bovine serum, 2 mM L-glutamine, and 0.1 mM nonessential amino acids (Invitrogen). Cells were maintained at 37°C and 5% CO₂. Cells were passaged with 0.05% trypsin-ethylene diamine tetraacetic acid. We cultured 3040-iPSCs on human embryonic stem cell-qualified Matrigel-coated plates (BD Biosciences, San Jose, CA) in mTeSR1 media (STEMCELL Technologies, Vancouver, Canada), with media changed daily.^{(20 (link))} Cells were passaged using gentle cell dissociation buffer (STEMCELL Technologies) with 10 mM ROCK inhibitor (Selleck Chemical, Houston, TX) and maintained at 37°C and 5% CO₂.^{(20 (link))} The human embryonic stem cell line H1 (WiCell, Madison, WI) was cultured as the iPSCs.