STAR Protocols (Sep 2023)
Enrichment of carcinogen-driven “mitochondria-primed” human skin stem cells and their identification using single-cell analyses
Abstract
Summary: Previous work has shown that mitochondria play a critical role in priming stem cells to self-renew and proliferate. Here, we describe a protocol for enriching and identifying the mitochondria-primed stem cells (mpSCs) for their characterization and applications. We describe steps for enriching mpSCs with the environmental carcinogen 2,3,7,8-tetrachlorodibenzo-p-dioxin in a skin keratinocyte lineage and for identifying mpSCs using single-cell transcriptomics and single-cell microscopy analyses of expression of relevant stem cell markers.For complete details on the use and execution of this protocol, please refer to Spurlock et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.