Human ES cell line WIBR1/2/3 and hES-Rett were previously described 26,44 and cultured in 5% O2 on mitomycin C-inactivated mouse embryonic fibroblasts (MEFs) in hESC medium, containing DMEM/F12 (Thermo), 15% fetal bovine serum (Hyclone), 5% knockout serum replacement (Thermo), 1% non-essential amino acids (Invitrogen), 1mM glutamine (Thermo), 0.1mM β-mercaptoethanol (Sigma) and 4ng/ml bFGF (Thermo). Cultures were passaged manually or with 1mg/ml collagenase type IV (Thermo) every 5–7 days. iPS-wt1/2/3, iPS-fAD2, iPS-AMN1/2/3 and iPS-ALD1/2/3/4 were reprogrammed from patient fibroblasts using the constitutive excisable STEMCCA lentivirus (OSKM) and maintained in the same conditions as WIBR1/2/3. iPS-wt4/5, iPS-fAD1/3 were reprogrammed using non-integrative Sendai virus (OSKML), and maintained on inactivated mouse embryonic fibroblasts in serum-free hES medium containing DMEM/F12 (Thermo), 20% knockout serum replacement (Thermo), 1% non-essential amino acids (Thermo), 1mM Glutamax (Thermo), 0.1mM β-mercaptoethanol (Sigma) and 12ng/ml bFGF (Thermo). hES-WIBR3 is a female line, all other lines are male. All parental lines were maintained for over 50 passages, verified for stable expression of Oct3/4, Nanog and TRA1-60, and routinely tested for mycoplasma negativity.
