Critical intermediates

pHAES-WCB (Starting material) -> pHAES10 -> EB96 -> HAES

1. pHEAS10: Propagation with MEF in KSR (LifeTec KSR) + KO-DMEM + FGF2.

2. EB96: Period of EB formation

A. 4 days in XF(-) [KO-DMEM, 20% Xeno-free SR] in 96-well dishes.

3. qHAES: Period of adhesion.

A. 28 to 35 days in XF32 [KO-DMEM, 15% Xeno-free SR] + (H + IGF1 + FGF2).

Critical intermediate

Critical parameter

Critical procedure

pHAES-WCB (Starting material) -> pHAES10 -> EB96 -> HAES

Sampling

pHAES10: 10-cm dish cultivation just before EB formation

EB96: EBs just before passage into T25 flasks.