Single cell sequencing of human primary LCs

Langerhans cells (LC) coordinate immune homeostasis at the human epidermis, proficent in initiating tolerogenic immune responses. To investigate the transcriptomic mediators of human LC tolerogenic immune responses, we performed single cell RNA-sequencing of steady state LC extracted via Liberase TM digestion and migratory LCs extracted from epidermal sheets. Steady state and Migrated human LC were subjected to single cell RNA-sequencing following encapsulation with Drop-seq. Following digest with dispase overnight at +4C, Steady state LC were extracted from epidermis by 90 minute Liberase TM digestion +37C and Migrated LC were extracted from 48 hour culture of epidermis sheets.