title: GSM7112592: Stromal, dermis, replicate 2; Mus musculus; RNA-Seq
library_construction_protocol: Back skins from C57BL/6 female adult mice were harvested after shaving with an electric clipper. Subcutaneous fat was removed from hypodermis and hypodermal layers were physically isolated from the dermis with forceps. Dermis was isolated from epidermis after 45min of an enzymatic treatment with 0.15% Tripsin/0.75mM EDTA. Isolated dermis and hypodermis were each minced with scissors and digested for 2H at 37°C in RPMI supplemented with 5% FCS and containing 0.25mg/mL of liberase T-Flex and 1ug/mL of DNAse I. Cells were isolated from each layer and immunostained with anti-EpCAM and anti-CD45 antibodies as well as stained with propidium iodide (PI) before being FACS sorted as live leucocytes (PI–, EpCAM–, CD45+) or live stromal cells (PI–, EpCAM–, CD45–). Cells were then washed, counted and resuspended into PBS containing 0.01% bovine serum albumin before being used for single-cell RNA-sequencing. Library construction was performed in accordance with 10X Genomics protocol (10X Chromium 3' v2 kit).