GSM5121234: Limbal Epithelial SCs- WT; Mus musculus; RNA-Seq
Ten eyes of 2.5 months old Krt15-GFP mice were enucleated, the limbus (with marginal conjunctiva and peripheral cornea) was dissected (~0.5 mm), tissues were pooled and incubated in trypsin (X10 Biological Industries) for 10 min 37°C 300 μl. Supernatant was collected into 10 ml (RPMI (Biological Industries) 10% chelated fetal calf serum). Trypsinization was repeated for 10 cycles adding fresh trypsin in each cycle. Cell suspension was centrifuged (8 min at 300g), re-suspended and filtered using Cell strainer (Danyel Biotech) to achieve 1200 cells/μL. An RNA library was produced according to the 10x Genomics protocol (Chromium Single Cell 3' Library & Gel Bead Kit v2, PN-120237) using 18000 input cells. scRNA-seq