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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| Format: | Article |
| Language: | English |
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MDPI AG
2022-10-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/11/21/3356 |
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| author | Daria Kostina Arseniy Lobov Polina Klausen Vitaly Karelkin Rashid Tikhilov Svetlana Bozhkova Andrey Sereda Nadezhda Ryumina Natella Enukashvily Anna Malashicheva |
| author_facet | Daria Kostina Arseniy Lobov Polina Klausen Vitaly Karelkin Rashid Tikhilov Svetlana Bozhkova Andrey Sereda Nadezhda Ryumina Natella Enukashvily Anna Malashicheva |
| author_sort | Daria Kostina |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T19:11:13Z |
| format | Article |
| id | doaj.art-3f7d2fc5f6c24b4c9c4a91cd9b51b0af |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T19:11:13Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-3f7d2fc5f6c24b4c9c4a91cd9b51b0af2023-11-24T04:07:23ZengMDPI AGCells2073-44092022-10-011121335610.3390/cells11213356Isolation of Human Osteoblast Cells Capable for Mineralization and Synthetizing Bone-Related Proteins In Vitro from Adult BoneDaria Kostina0Arseniy Lobov1Polina Klausen2Vitaly Karelkin3Rashid Tikhilov4Svetlana Bozhkova5Andrey Sereda6Nadezhda Ryumina7Natella Enukashvily8Anna Malashicheva9Institute of Cytology, Russian Academy of Science, Tikhoretskiy Prospect 4, 194064 St. Petersburg, RussiaInstitute of Cytology, Russian Academy of Science, Tikhoretskiy Prospect 4, 194064 St. Petersburg, RussiaInstitute of Cytology, Russian Academy of Science, Tikhoretskiy Prospect 4, 194064 St. Petersburg, RussiaVreden National Medical Research Center of Traumatology and Orthopedics, National Medical Research Center, Street Academician Baykova, 8, 195427 St. Petersburg, RussiaVreden National Medical Research Center of Traumatology and Orthopedics, National Medical Research Center, Street Academician Baykova, 8, 195427 St. Petersburg, RussiaVreden National Medical Research Center of Traumatology and Orthopedics, National Medical Research Center, Street Academician Baykova, 8, 195427 St. Petersburg, RussiaVreden National Medical Research Center of Traumatology and Orthopedics, National Medical Research Center, Street Academician Baykova, 8, 195427 St. Petersburg, RussiaInstitute of Cytology, Russian Academy of Science, Tikhoretskiy Prospect 4, 194064 St. Petersburg, RussiaInstitute of Cytology, Russian Academy of Science, Tikhoretskiy Prospect 4, 194064 St. Petersburg, RussiaInstitute of Cytology, Russian Academy of Science, Tikhoretskiy Prospect 4, 194064 St. Petersburg, RussiaThe 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.https://www.mdpi.com/2073-4409/11/21/3356osteoblastsosteogenic differentiationprimary cell culture |
| spellingShingle | Daria Kostina Arseniy Lobov Polina Klausen Vitaly Karelkin Rashid Tikhilov Svetlana Bozhkova Andrey Sereda Nadezhda Ryumina Natella Enukashvily Anna Malashicheva Isolation of Human Osteoblast Cells Capable for Mineralization and Synthetizing Bone-Related Proteins In Vitro from Adult Bone Cells osteoblasts osteogenic differentiation primary cell culture |
| 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 | osteoblasts osteogenic differentiation primary cell culture |
| url | https://www.mdpi.com/2073-4409/11/21/3356 |
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