GSM8994785: 3w_2; Mus musculus; RNA-Seq
For single-cell RNA sequencing (scRNA-seq) analysis, 3- and 8-week-old C57BL/6J wild-type mice were used. The diaphysis was trimmed approximately 2 mm distal to the growth plate, and the metaphyseal (MP) regions of distal femurs and proximal tibias were collected in digestion buffer. Following bone dissociation as described above, FACS was performed using a FACS Melody system (BD Biosciences). CD31/CD45/TER119-negative cells were sorted after dead cells were excluded using 7-AAD (1:200; BioLegend, 420403). Sorted cells were immediately collected into an ice-cold cell suspension buffer and loaded onto a Chromium Controller (10X Genomics). Single-cell libraries were prepared using the Chromium Next GEM Single Cell 3′ Reagent Kits v3.1 (10X Genomics).