H1 hESCs (male embryo), control hiPSC lines hVS-88 (74 days old male), hVS-60 (70 year old male) and hVS-421 (19 year old male) henceforth referred to as Line A, B, C and D respectively, were cultured with a feeder independent method on Geltrex in Essential 8 medium (GIBCO). The human ESC line HO1 was obtained from WiCell. The hiPSC control lines (hvs-88 and 60) were generated via reprogramming fibroblasts from two healthy individuals (fibroblasts were derived from anonymous, non-identifiable donors and therefore exempt from IRB approval). One hNES cell line was generated [25] from each stem cell line A, B and C. Repetitive differentiation experiments performed from one hNES culture are referred to as B1, B2, B3, etc.
