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Isolation and in vitro characterization of murine young-adult long bone skeletal progenitors

Skeletal stem and progenitor cells (SSPCs) constitute a reservoir of bone-forming cells necessary for bone development, modeling and remodeling, as well as for fracture healing. Recent advances in tools to identify and isolate SSPCs have revealed that cells with multipotent properties are present no...

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Autores principales: Loopmans, Shauni, Stockmans, Ingrid, Carmeliet, Geert, Stegen, Steve
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9376626/
https://www.ncbi.nlm.nih.gov/pubmed/35979436
http://dx.doi.org/10.3389/fendo.2022.930358
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author Loopmans, Shauni
Stockmans, Ingrid
Carmeliet, Geert
Stegen, Steve
author_facet Loopmans, Shauni
Stockmans, Ingrid
Carmeliet, Geert
Stegen, Steve
author_sort Loopmans, Shauni
collection PubMed
description Skeletal stem and progenitor cells (SSPCs) constitute a reservoir of bone-forming cells necessary for bone development, modeling and remodeling, as well as for fracture healing. Recent advances in tools to identify and isolate SSPCs have revealed that cells with multipotent properties are present not only in neonatal bone, but also in adult bone marrow and periosteum. The long bone metaphysis and endosteum have been proposed as an additional SSPC niche, although in vitro approaches to study their cellular and molecular characteristics are still limited. Here, we describe a comprehensive procedure to isolate and culture SSPCs derived from the metaphysis and endosteum of young-adult mice. Based on flow cytometry analysis of known SSPC markers, we found the presence of putative multipotent SSPCs, similar to neonatal bone tissue. In vitro, metaphyseal/endosteal SSPCs possess self-renewing capacity, and their multipotency is underscored by the ability to differentiate into the osteogenic and adipogenic lineage, while chondrogenic potential is limited. Expansion of metaphyseal/endosteal SSPCs under low oxygen conditions increases their proliferation capacity, while progenitor properties are maintained, likely reflecting their hypoxic niche in vivo. Collectively, we propose a validated isolation and culture protocol to study metaphyseal/endosteal SSPC biology in vitro.
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spelling pubmed-93766262022-08-16 Isolation and in vitro characterization of murine young-adult long bone skeletal progenitors Loopmans, Shauni Stockmans, Ingrid Carmeliet, Geert Stegen, Steve Front Endocrinol (Lausanne) Endocrinology Skeletal stem and progenitor cells (SSPCs) constitute a reservoir of bone-forming cells necessary for bone development, modeling and remodeling, as well as for fracture healing. Recent advances in tools to identify and isolate SSPCs have revealed that cells with multipotent properties are present not only in neonatal bone, but also in adult bone marrow and periosteum. The long bone metaphysis and endosteum have been proposed as an additional SSPC niche, although in vitro approaches to study their cellular and molecular characteristics are still limited. Here, we describe a comprehensive procedure to isolate and culture SSPCs derived from the metaphysis and endosteum of young-adult mice. Based on flow cytometry analysis of known SSPC markers, we found the presence of putative multipotent SSPCs, similar to neonatal bone tissue. In vitro, metaphyseal/endosteal SSPCs possess self-renewing capacity, and their multipotency is underscored by the ability to differentiate into the osteogenic and adipogenic lineage, while chondrogenic potential is limited. Expansion of metaphyseal/endosteal SSPCs under low oxygen conditions increases their proliferation capacity, while progenitor properties are maintained, likely reflecting their hypoxic niche in vivo. Collectively, we propose a validated isolation and culture protocol to study metaphyseal/endosteal SSPC biology in vitro. Frontiers Media S.A. 2022-08-01 /pmc/articles/PMC9376626/ /pubmed/35979436 http://dx.doi.org/10.3389/fendo.2022.930358 Text en Copyright © 2022 Loopmans, Stockmans, Carmeliet and Stegen https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Endocrinology
Loopmans, Shauni
Stockmans, Ingrid
Carmeliet, Geert
Stegen, Steve
Isolation and in vitro characterization of murine young-adult long bone skeletal progenitors
title Isolation and in vitro characterization of murine young-adult long bone skeletal progenitors
title_full Isolation and in vitro characterization of murine young-adult long bone skeletal progenitors
title_fullStr Isolation and in vitro characterization of murine young-adult long bone skeletal progenitors
title_full_unstemmed Isolation and in vitro characterization of murine young-adult long bone skeletal progenitors
title_short Isolation and in vitro characterization of murine young-adult long bone skeletal progenitors
title_sort isolation and in vitro characterization of murine young-adult long bone skeletal progenitors
topic Endocrinology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9376626/
https://www.ncbi.nlm.nih.gov/pubmed/35979436
http://dx.doi.org/10.3389/fendo.2022.930358
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