Transcriptomic single-cell analysis of murine neonatal growth plate with conditional G6PDH deletion
Transcriptomic single-cell analysis of murine neonatal growth plate with conditional G6PDH deletion
Growth plates were isolated from the distal femur and proximal tibia of 3-day-old mice (male wild-type: n=1; male G6PDH-null: n=1; female wild-type: n=1; female G6PDH-null: n=1) and the perichondrium was removed. To obtain a single cell suspension, the growth plates were digested with 5 mg/ml collagenase type II in minimum essential medium alpha GLUTAMAXTM supplemented with 100 units/ml penicillin, 50 µg/ml streptomycin and 10% FBS (all from Gibco, ThermoFisher Scientific) for 1.5 hours at 37°C. The cell suspension was passed through a 40 µm nylon mesh, centrifuged at 300g for 7 minutes at room temperature and resuspended in phosphate-buffered saline supplemented with bovine serum albumin.  Single cell suspensions were converted to barcoded scRNA-seq libraries using the Chromium Single Cell 3' Reagent Kits v3 (10x Genomics), aiming for 5,000 cells per library.