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

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 tran...

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Detalles Bibliográficos
Autores principales: Kilpatrick, Alastair Morris, Marques, Madalena, Gomez-Salazar, Mario, Stefancova, Dorota, Sá da Bandeira, Diana, Crisan, Mihaela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9846014/
https://www.ncbi.nlm.nih.gov/pubmed/36640365
http://dx.doi.org/10.1016/j.xpro.2022.102016
Descripción
Sumario: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)

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