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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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
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author Kilpatrick, Alastair Morris
Marques, Madalena
Gomez-Salazar, Mario
Stefancova, Dorota
Sá da Bandeira, Diana
Crisan, Mihaela
author_facet Kilpatrick, Alastair Morris
Marques, Madalena
Gomez-Salazar, Mario
Stefancova, Dorota
Sá da Bandeira, Diana
Crisan, Mihaela
author_sort Kilpatrick, Alastair Morris
collection PubMed
description 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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spelling pubmed-98460142023-01-19 Protocol to analyze and validate transcriptomic changes in PDGFRβ-KO mesenchymal stem cell osteogenic potential in the mouse embryo Kilpatrick, Alastair Morris Marques, Madalena Gomez-Salazar, Mario Stefancova, Dorota Sá da Bandeira, Diana Crisan, Mihaela STAR Protoc Protocol 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) Elsevier 2023-01-12 /pmc/articles/PMC9846014/ /pubmed/36640365 http://dx.doi.org/10.1016/j.xpro.2022.102016 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Kilpatrick, Alastair Morris
Marques, Madalena
Gomez-Salazar, Mario
Stefancova, Dorota
Sá da Bandeira, Diana
Crisan, Mihaela
Protocol to analyze and validate transcriptomic changes in PDGFRβ-KO mesenchymal stem cell osteogenic potential in the mouse embryo
title Protocol to analyze and validate transcriptomic changes in PDGFRβ-KO mesenchymal stem cell osteogenic potential in the mouse embryo
title_full Protocol to analyze and validate transcriptomic changes in PDGFRβ-KO mesenchymal stem cell osteogenic potential in the mouse embryo
title_fullStr Protocol to analyze and validate transcriptomic changes in PDGFRβ-KO mesenchymal stem cell osteogenic potential in the mouse embryo
title_full_unstemmed Protocol to analyze and validate transcriptomic changes in PDGFRβ-KO mesenchymal stem cell osteogenic potential in the mouse embryo
title_short Protocol to analyze and validate transcriptomic changes in PDGFRβ-KO mesenchymal stem cell osteogenic potential in the mouse embryo
title_sort protocol to analyze and validate transcriptomic changes in pdgfrβ-ko mesenchymal stem cell osteogenic potential in the mouse embryo
topic Protocol
url 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
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