Generation of two multipotent mesenchymal progenitor cell lines capable of osteogenic, mature osteocyte, adipogenic, and chondrogenic differentiation
Abstract Mesenchymal progenitors differentiate into several tissues including bone, cartilage, and adipose. Targeting these cells in vivo is challenging, making mesenchymal progenitor cell lines valuable tools to study tissue development. Mesenchymal stem cells (MSCs) can be isolated from humans and...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2021-11-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-02060-1 |
| _version_ | 1798034553572425728 |
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| author | Matthew Prideaux Christian S. Wright Megan L. Noonan Xin Yi Erica L. Clinkenbeard Elsa Mevel Jonathan A. Wheeler Sharon Byers Asiri R. Wijenayaka Stan Gronthos Uma Sankar Kenneth E. White Gerald J. Atkins William R. Thompson |
| author_facet | Matthew Prideaux Christian S. Wright Megan L. Noonan Xin Yi Erica L. Clinkenbeard Elsa Mevel Jonathan A. Wheeler Sharon Byers Asiri R. Wijenayaka Stan Gronthos Uma Sankar Kenneth E. White Gerald J. Atkins William R. Thompson |
| author_sort | Matthew Prideaux |
| collection | DOAJ |
| description | Abstract Mesenchymal progenitors differentiate into several tissues including bone, cartilage, and adipose. Targeting these cells in vivo is challenging, making mesenchymal progenitor cell lines valuable tools to study tissue development. Mesenchymal stem cells (MSCs) can be isolated from humans and animals; however, obtaining homogenous, responsive cells in a reproducible fashion is challenging. As such, we developed two mesenchymal progenitor cell (MPC) lines, MPC1 and MPC2, generated from bone marrow of male C57BL/6 mice. These cells were immortalized using the temperature sensitive large T-antigen, allowing for thermal control of proliferation and differentiation. Both MPC1 and MPC2 cells are capable of osteogenic, adipogenic, and chondrogenic differentiation. Under osteogenic conditions, both lines formed mineralized nodules, and stained for alizarin red and alkaline phosphatase, while expressing osteogenic genes including Sost, Fgf23, and Dmp1. Sost and Dmp1 mRNA levels were drastically reduced with addition of parathyroid hormone, thus recapitulating in vivo responses. MPC cells secreted intact (iFGF23) and C-terminal (cFGF23) forms of the endocrine hormone FGF23, which was upregulated by 1,25 dihydroxy vitamin D (1,25D). Both lines also rapidly entered the adipogenic lineage, expressing adipose markers after 4 days in adipogenic media. MPC cells were also capable of chondrogenic differentiation, displaying increased expression of cartilaginous genes including aggrecan, Sox9, and Comp. With the ability to differentiate into multiple mesenchymal lineages and mimic in vivo responses of key regulatory genes/proteins, MPC cells are a valuable model to study factors that regulate mesenchymal lineage allocation as well as the mechanisms that dictate transcription, protein modification, and secretion of these factors. |
| first_indexed | 2024-04-11T20:44:55Z |
| format | Article |
| id | doaj.art-f7398838cd214c8aba8d0dac83a09d11 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-11T20:44:55Z |
| publishDate | 2021-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-f7398838cd214c8aba8d0dac83a09d112022-12-22T04:04:04ZengNature PortfolioScientific Reports2045-23222021-11-0111111610.1038/s41598-021-02060-1Generation of two multipotent mesenchymal progenitor cell lines capable of osteogenic, mature osteocyte, adipogenic, and chondrogenic differentiationMatthew Prideaux0Christian S. Wright1Megan L. Noonan2Xin Yi3Erica L. Clinkenbeard4Elsa Mevel5Jonathan A. Wheeler6Sharon Byers7Asiri R. Wijenayaka8Stan Gronthos9Uma Sankar10Kenneth E. White11Gerald J. Atkins12William R. Thompson13Indiana Center for Musculoskeletal Health, Indiana UniversityIndiana Center for Musculoskeletal Health, Indiana UniversityIndiana Center for Musculoskeletal Health, Indiana UniversityDepartment of Physical Therapy, School of Health and Human Sciences, Indiana UniversityIndiana Center for Musculoskeletal Health, Indiana UniversityDepartment of Anatomy, Cell Biology, and Physiology, Indiana UniversityDepartment of Medical and Molecular Genetics, Indiana UniversityCentre for Orthopaedic and Trauma Research, Faculty of Health and Medical Sciences, University of AdelaideCentre for Orthopaedic and Trauma Research, Faculty of Health and Medical Sciences, University of AdelaideMesenchymal Stem Cell Laboratory, Faculty of Health and Medical Sciences, University of AdelaideIndiana Center for Musculoskeletal Health, Indiana UniversityIndiana Center for Musculoskeletal Health, Indiana UniversityCentre for Orthopaedic and Trauma Research, Faculty of Health and Medical Sciences, University of AdelaideIndiana Center for Musculoskeletal Health, Indiana UniversityAbstract Mesenchymal progenitors differentiate into several tissues including bone, cartilage, and adipose. Targeting these cells in vivo is challenging, making mesenchymal progenitor cell lines valuable tools to study tissue development. Mesenchymal stem cells (MSCs) can be isolated from humans and animals; however, obtaining homogenous, responsive cells in a reproducible fashion is challenging. As such, we developed two mesenchymal progenitor cell (MPC) lines, MPC1 and MPC2, generated from bone marrow of male C57BL/6 mice. These cells were immortalized using the temperature sensitive large T-antigen, allowing for thermal control of proliferation and differentiation. Both MPC1 and MPC2 cells are capable of osteogenic, adipogenic, and chondrogenic differentiation. Under osteogenic conditions, both lines formed mineralized nodules, and stained for alizarin red and alkaline phosphatase, while expressing osteogenic genes including Sost, Fgf23, and Dmp1. Sost and Dmp1 mRNA levels were drastically reduced with addition of parathyroid hormone, thus recapitulating in vivo responses. MPC cells secreted intact (iFGF23) and C-terminal (cFGF23) forms of the endocrine hormone FGF23, which was upregulated by 1,25 dihydroxy vitamin D (1,25D). Both lines also rapidly entered the adipogenic lineage, expressing adipose markers after 4 days in adipogenic media. MPC cells were also capable of chondrogenic differentiation, displaying increased expression of cartilaginous genes including aggrecan, Sox9, and Comp. With the ability to differentiate into multiple mesenchymal lineages and mimic in vivo responses of key regulatory genes/proteins, MPC cells are a valuable model to study factors that regulate mesenchymal lineage allocation as well as the mechanisms that dictate transcription, protein modification, and secretion of these factors.https://doi.org/10.1038/s41598-021-02060-1 |
| spellingShingle | Matthew Prideaux Christian S. Wright Megan L. Noonan Xin Yi Erica L. Clinkenbeard Elsa Mevel Jonathan A. Wheeler Sharon Byers Asiri R. Wijenayaka Stan Gronthos Uma Sankar Kenneth E. White Gerald J. Atkins William R. Thompson Generation of two multipotent mesenchymal progenitor cell lines capable of osteogenic, mature osteocyte, adipogenic, and chondrogenic differentiation Scientific Reports |
| title | Generation of two multipotent mesenchymal progenitor cell lines capable of osteogenic, mature osteocyte, adipogenic, and chondrogenic differentiation |
| title_full | Generation of two multipotent mesenchymal progenitor cell lines capable of osteogenic, mature osteocyte, adipogenic, and chondrogenic differentiation |
| title_fullStr | Generation of two multipotent mesenchymal progenitor cell lines capable of osteogenic, mature osteocyte, adipogenic, and chondrogenic differentiation |
| title_full_unstemmed | Generation of two multipotent mesenchymal progenitor cell lines capable of osteogenic, mature osteocyte, adipogenic, and chondrogenic differentiation |
| title_short | Generation of two multipotent mesenchymal progenitor cell lines capable of osteogenic, mature osteocyte, adipogenic, and chondrogenic differentiation |
| title_sort | generation of two multipotent mesenchymal progenitor cell lines capable of osteogenic mature osteocyte adipogenic and chondrogenic differentiation |
| url | https://doi.org/10.1038/s41598-021-02060-1 |
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