Characterization of human iPSC-derived ocular surface epithelial cell sheets depending on PAX6 gene expression levels
Characterization of human iPSC-derived ocular surface epithelial cell sheets depending on PAX6 gene expression levels
Sorted hiPSCs were plated onto iMatrix511silk-coated (0.5 μg/cm2) cell culture inserts. The cells were cultured in OEM (DMEM/F12 containing 10 μM Y-27632, 2% B27 supplement, 1% penicillin-streptomycin solution, and 10ng/m of KGF) at 37 °C until confluence (4 weeks).  Total RNA was purified using an RNeasy Micro Kit with RNase-free DNase (both from Qiagen, Hilden, Germany). RNA-seq libraries were prepared using the TruSeq Stranded mRNA Library Prep Kit (Illumina), and the quality and quantity of the amplicons were assessed at all steps through capillary electrophoresis (Agilent Bioanalyzer and TapeStation; Agilent, Santa Clara, CA, USA).