Protocol to analyze and validate transcriptomic changes in PDGFRO-KO mesenchymal stem cell osteogenic potential in the mouse embryo

Mesenchymal stem/stromal cells (MSCs) can differentiate into osteoblasts under appropriate conditions. PDGFRO signaling controls MSC osteogenic potential both transcriptomically and in culture. Here, we present a "computer to the bench"protocol to analyze changes in MSC osteogenic potential at transcrip-tomic and cellular level in the absence of PDGFRO. We detail the preparation of cells from mouse embryos, the analysis of transcriptomic changes from sin-gle-cell RNA-sequencing data, the procedure for MSC derivation and culture, and an osteogenic assay for functional validation. For complete details on the use and execution of this protocol, please refer to Sada Bandeira et al. (2022).1