The Simplest Protocol for Rapid and Long-Term Culture of Primary Epidermal Keratinocytes from Human and Mouse

The document introduced a protocol for the long-term culture of primary epidermal keratinocytes from humans and mice, focusing on p63+ mouse epidermal keratinocytes. The protocol involved the use of low calcium media and the pharmacological inhibition of TGF-β signaling, which increased the proliferative capacity of these cells without requiring progenitor cell purification or feeder cell layers. This method allowed for the formation of large clones in co-culture and enhanced the growth of human keratinocytes with various feeder cells. The approach aimed to facilitate the use of mouse models and expedite the preparation of customized skin grafts.