3D and Air Liquid Interface culture of human iPSC-derived alveoar epithelial type I-like cells (iAT1s) [NGAT LDCI]

The goal of this study is to understand transcriptomic differences of human iPSC derived alveolar epithelial type I-like cells cultured in 3D and at Air Liquid Interface (ALI). Overall design: BU3 NGAT iPSCs were differentiated into iPSC derived alveolar epithelial type 2 cells (iAT2s). Cells were either maintained as iAT2s in 3D Matrigel in CKDCI medium or differentiated into iAT1s using LDCI medium. ALI versions of iAT1 cultures were prepared as follows: to prepare “iAT1 ALI p0” cultures, first a single cell suspension of iAT2s was passaged onto 6.5mm transwell inserts (Corning) coated with hESC qualified Matrigel (Corning, 8774552), while switching CKDCI medium to LDCI at the time of plating. To prepare “iAT1 ALI p1” cultures, iAT2s in 3D Matrigel culture were first switched to LDCI medium in 3D for 9 days before being passaged as single cells onto Matrigel coated transwells for continued LDCI culture. All ALI cultures were continued for 10 days after transwell plating.