Novel Protocol for the In Vitro Maintenance and Expansion of Adult Epidermal LGR5+ Stem Cells

The study developed a novel culture method for maintaining and expanding LGR5+ epidermal adult stem cells using a transgenic porcine model with a GFP tag. The research compared the effectiveness of porcine fetal fibroblasts (PFF) and mouse embryonic fibroblasts (MEF) as feeder layers, testing three different media conditions. Results indicated that MEF feeder layers supported a higher growth rate and maintained the LGR5+ lineage more effectively than PFFs. Specifically, media containing 5-azacytidine and CHIR99021 were crucial for maintaining the LGR5+ lineage. The study suggested that this protocol could be used to create three-dimensional skin growth in vitro, potentially improving grafts for burn victims.