Generation of Human Induced Pluripotent Stem Cell-Derived Planar Hair-Bearing Skin Organoids Using an Air-Liquid Interface Culture System

The document outlines a protocol for generating human induced pluripotent stem cell-derived skin organoids (SOs) with hair follicles using an air-liquid interface (ALI) culture system. Initially developed in an inverted cystic form, these SOs are transformed into a planar configuration that better mimics human skin. The protocol includes checkpoints for assessing organoid development, steps for flattening cystic SOs, and optimal ALI culture conditions. By Day 27, the organoids exhibit a pluristratified pigmented epithelium with hair follicles and sebaceous gland-like structures. This method provides a physiologically relevant in vitro model for skin research, applicable in studies of pathogen infection, UV-induced damage, and drug responses.