Effect of externally applied concave and convex curvature for 24 hours with continued duam SMAD inhibition (dSMADi) on human embryonic cell line H9 after 48 hours of dSMADi induction to attain early neuroepithelial features
To investigate the transcriptional change induced by externally applied concave and convex curvature, the former of which displayed tissue-level morphological change, human embryonic cell line H9 maintained for pluripotency with mTeSR1 for 72 hours and then induced to have early neuroepithelial features with N2B27 media containing 10 µM of SB431542 and 0.1 µM of LDN (dual SMAD inhibition) for 48 hours. Then along with the continued dual SMAD inhibition, plate curving cell culture of either concave or convex conformation for 24 hours was applied on the cell culture, at the end of which time point cell pellet was collected for bulk RNA sequencing. Overall design: Triplicates of control (only without plate curving cell culture), concave curvature and convex curvature groups of human embryonic stem cell line-derived neuropithelium (hESC-NE) were prepared. Each two-member combination was compared to reveal differential expression.