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

Abstract

Summary: Mesenchymal stem/stromal cells (MSCs) can differentiate into osteoblasts under appropriate conditions. PDGFRβ 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 transcriptomic and cellular level in the absence of PDGFRβ. We detail the preparation of cells from mouse embryos, the analysis of transcriptomic changes from single-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 Sá da Bandeira et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Bioinformatics
• Cell Biology
• Single Cell
• Developmental Biology
• RNAseq
• Molecular Biology