GSM4191344: 161088C_D-4; Mus musculus; RNA-Seq
Human: Human EMPD (Patient ID; n=1) and normal skin (Patient ID; n=1; normal skin constitutes site adjacent to EMPD lesion). Samples were surgically excised and placed in tissue storage solution (MACS). Samples were processed for single cell isolation within 10 hours post-surgery. Excess subcutaneous adipose and dermal tissues were excised. Samples were cut into pieces with a maximum diameter of 4 mm. Skin specimens were then washed twice with cold 1X PBS. Single cells were isolated using the human epidermis isolation kit (MACS) as per manufacturer's instructions with minor modifications. Mouse: Skin epidermal cells were collected from DTG (n=1) and control (n=1) newborn mice (post-natal day 0.5) 5 days after Doxycycline treatment. Mice were euthanized, and consecutively immersed in 70% ethanol and cold 1X PBS twice for 3 minutes. Fore and hind limbs and tail were cut off and skin was peeled along the ventral midline axis with blunt tweezers. Subcutaneous adipose tissue was micro-dissected. 12 mm skin biopsies were laid on EpiLife CF basal media (STEM CELL) droplets (300 μL) on a 12-well plate with skin dermis side down. 2 ml (2.5 U/mL) Dispase solution (STEM CELL)  was added to skin and incubated at 4℃ for 12 hrs. Epidermis was separated from dermis with two sharp tweezers and placed on Accutase (ThermoFisher) droplets (500 μL) for 30 min (dermis side down) at RT to create single cell suspensions. Single cells were captured using the Chromium Platform (10X Genomics) and libraries were generated using Single Cell 3' v2 chemistry (mouse samples) and Single Cell 3' v3 chemistry (human samples). Cell capture, library preparation, quality control, and sequencing was performed at Beijing CapitalBio Technology (Beijing, China). Single Cell 3' v2 chemistry (mouse samples) - 10X Genomics
GEO Accession
GSM4191344