Single-Cell RNA Sequencing of Calvarial and Long-Bone Endocortical Cells

Ugur M Ayturk, Joseph P Scollan, Didem Goz Ayturk, Eun Sung Suh, Alexander Vesprey, Christina M Jacobsen, Paola Divieti Pajevic, Matthew L Warman

DOI:10.1002/jbmr.4052

We obtained primary cells by enzymatically digesting P5 mouse calvariae. Part of the cells were processed for single cell RNA-seq immediately. The remaining cells were cultured in 6-well plates till they reached confluence at 5 days. Cultured cells were then trypsin-treated, transferred to collagen-coated plates and incubated in osteogenic medium (containing ascorbic acid and beta-glycerophosphate) for an additional 7-days. Cells were then collected and subjected to single cell RNA-seq.