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Isolation of Human Osteoblast Cells Capable for Mineralization and Synthetizing Bone-Related Proteins In Vitro from Adult Bone

The culture of osteoblasts (OB) of human origin is a useful experimental model in studying bone biology, osteogenic differentiation, functions of bone proteins, oncological processes in bone tissue, testing drugs against bone desires, and many other fields. The purpose of the present study is to sha...

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Autores principales: Kostina, Daria, Lobov, Arseniy, Klausen, Polina, Karelkin, Vitaly, Tikhilov, Rashid, Bozhkova, Svetlana, Sereda, Andrey, Ryumina, Nadezhda, Enukashvily, Natella, Malashicheva, Anna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9654174/
https://www.ncbi.nlm.nih.gov/pubmed/36359752
http://dx.doi.org/10.3390/cells11213356
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author Kostina, Daria
Lobov, Arseniy
Klausen, Polina
Karelkin, Vitaly
Tikhilov, Rashid
Bozhkova, Svetlana
Sereda, Andrey
Ryumina, Nadezhda
Enukashvily, Natella
Malashicheva, Anna
author_facet Kostina, Daria
Lobov, Arseniy
Klausen, Polina
Karelkin, Vitaly
Tikhilov, Rashid
Bozhkova, Svetlana
Sereda, Andrey
Ryumina, Nadezhda
Enukashvily, Natella
Malashicheva, Anna
author_sort Kostina, Daria
collection PubMed
description The culture of osteoblasts (OB) of human origin is a useful experimental model in studying bone biology, osteogenic differentiation, functions of bone proteins, oncological processes in bone tissue, testing drugs against bone desires, and many other fields. The purpose of the present study is to share a workflow that has established the conditions to efficiently isolate and grow OB cells obtained from surgically removed bones from human donors. The protocol described here also shows how to determine cell phenotype. Here we provide characteristics of cells isolated by this protocol that might help researchers to decide if such OB are suitable for the purposes of their study. Osteoblasts isolated from collagenase-treated explants of adult bones are able to proliferate and keep their phenotype in culture. OB cells have high synthetic properties. They express osteomarkers, such as RUNX2, osteocalcin, BMP2, and osteopontin both in control conditions and in an osteogenic medium that could be estimated by qPCR and immunocytochemical staining and by Western blotting. Induction of osteogenic differentiation does not dramatically influence the synthetic properties of OB cells, while the cells gain the ability to extracellular mineralization only in an osteogenic medium.
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spelling pubmed-96541742022-11-15 Isolation of Human Osteoblast Cells Capable for Mineralization and Synthetizing Bone-Related Proteins In Vitro from Adult Bone Kostina, Daria Lobov, Arseniy Klausen, Polina Karelkin, Vitaly Tikhilov, Rashid Bozhkova, Svetlana Sereda, Andrey Ryumina, Nadezhda Enukashvily, Natella Malashicheva, Anna Cells Article The culture of osteoblasts (OB) of human origin is a useful experimental model in studying bone biology, osteogenic differentiation, functions of bone proteins, oncological processes in bone tissue, testing drugs against bone desires, and many other fields. The purpose of the present study is to share a workflow that has established the conditions to efficiently isolate and grow OB cells obtained from surgically removed bones from human donors. The protocol described here also shows how to determine cell phenotype. Here we provide characteristics of cells isolated by this protocol that might help researchers to decide if such OB are suitable for the purposes of their study. Osteoblasts isolated from collagenase-treated explants of adult bones are able to proliferate and keep their phenotype in culture. OB cells have high synthetic properties. They express osteomarkers, such as RUNX2, osteocalcin, BMP2, and osteopontin both in control conditions and in an osteogenic medium that could be estimated by qPCR and immunocytochemical staining and by Western blotting. Induction of osteogenic differentiation does not dramatically influence the synthetic properties of OB cells, while the cells gain the ability to extracellular mineralization only in an osteogenic medium. MDPI 2022-10-24 /pmc/articles/PMC9654174/ /pubmed/36359752 http://dx.doi.org/10.3390/cells11213356 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kostina, Daria
Lobov, Arseniy
Klausen, Polina
Karelkin, Vitaly
Tikhilov, Rashid
Bozhkova, Svetlana
Sereda, Andrey
Ryumina, Nadezhda
Enukashvily, Natella
Malashicheva, Anna
Isolation of Human Osteoblast Cells Capable for Mineralization and Synthetizing Bone-Related Proteins In Vitro from Adult Bone
title Isolation of Human Osteoblast Cells Capable for Mineralization and Synthetizing Bone-Related Proteins In Vitro from Adult Bone
title_full Isolation of Human Osteoblast Cells Capable for Mineralization and Synthetizing Bone-Related Proteins In Vitro from Adult Bone
title_fullStr Isolation of Human Osteoblast Cells Capable for Mineralization and Synthetizing Bone-Related Proteins In Vitro from Adult Bone
title_full_unstemmed Isolation of Human Osteoblast Cells Capable for Mineralization and Synthetizing Bone-Related Proteins In Vitro from Adult Bone
title_short Isolation of Human Osteoblast Cells Capable for Mineralization and Synthetizing Bone-Related Proteins In Vitro from Adult Bone
title_sort isolation of human osteoblast cells capable for mineralization and synthetizing bone-related proteins in vitro from adult bone
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9654174/
https://www.ncbi.nlm.nih.gov/pubmed/36359752
http://dx.doi.org/10.3390/cells11213356
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