GSM4979487: Type_L_Diaphysis; Mus musculus; RNA-Seq
Femurs from wild-type mice were collected in PBS. Epiphysis, metaphysis and diaphysis were crushed separately with pestle in DMEM (+ Collagenase IV 10μl/ml, + Dispase 4μ/ml, + DNase 0.2μ/ml), and incubated 30min at 37°. The cells were dissociated to a single-cell suspension by filtering thorough 40μm nylon mesh. To eliminate red blood cells, we used RBC Lysis Buffer (pluriSelect 60-00050-11). Cells are FC-blocked and stained with FITC anti-mouse CD31 antibody (Biolegend 102506; 1:200), and APC anti mouse CD45 antibody (Biolegend 103112;1:200) for 40 min. Cell sorting was performed with Cell Sorter SH800S (SONY, Tokyo,Japan). Libraries were constructed using a Chromium Single Cell 3' v3.1 (10×Genomics, Pleasanton, CA) according to the manufacturer's instructions. single cell RNA-Seq