Human HF collection and organ culture

HF units from occipital region were obtained from hair transplant patients and whole scalp skin samples from face lift patients following ethical approval granted by East London and The City Research Ethics Committee 1 (REC Alpha 09/H0704/40) and written informed patient consent, adhering to the Declaration of Helsinki Principles. All samples, slides and biological material were tracked and stored according to the ‘Human Tissue Act’ guidelines. Anagen VI HFs were microdissected and cultured as previously described²⁷. ORS-KC were isolated as previously described⁵⁶ with a modification-freshly isolated ORS-KC were seeded in collagen I (Thermo Fisher, UK) coated flasks and cultured in Keratinocyte Serum Free Medium (Thermo Fisher, UK). Starvation was performed by incubation in SILAC RPMI 1640 Flex Medium (Thermo Fisher, UK). PYGL inhibitor (Cayman Chemicals, UK) and 2-NBDG (Thermo Fisher, UK) were diluted from stock DMSO solution in culture medium. Lactate (Sigma-Aldrich, UK) was diluted in culture media.