GSM8942337: BRONJ, CD45-; Mus musculus; RNA-Seq
Nine C57/B6 male mice were used to acquire single-cell suspensions each group. Mandibles were dissected with soft tissues removed. Then, mandible tissue was cut into small pieces and digested with 3 mg/mL collagenase type I (Gibco) and 4 mg/mL dispase II (Sigma) for 60 min at 37°C as previously described (Cui et al., 2020; Yamaza et al., 2011). Red blood cells were lysed with 1 mL Red Cell Lysis Solution (Biosharp). Cells were centrifuged and resuspended in 1% BSA solution. Then, mandible tissue was digested to acquire single cell suspension, which enriched with CD45- cells using Meiteni Mouse CD45 MicroBeads. The final concentration of cells was 1 x 10^6 cells per mL. Cellular suspensions were loaded into Chromium microfluidic chips and barcoded with a 10x Chromium Controller (10x Genomics). RNA from the barcoded cells was subsequently reverse-transcribed and sequencing libraries constructed with reagents from a Chromium Single Cell 3' v3 reagent kit (10x Genomics) and sequenced with the NovaSeq system (Illumina).