Murine Osteoblast and Osteoclast Co-culture on Demineralized Bone Paper for Bone Remodeling
Continuous and balanced bone remodeling is essential for maintaining mechanical integrity, mineral homeostasis, and hematopoiesis. Dysregulated bone metabolism develops pathological conditions, such as osteoporosis and bone metastasis. Functional and analytical recapitulation of bone remodeling in v...
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Bio-protocol LLC
2025-06-01
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| Online Access: | https://bio-protocol.org/en/bpdetail?id=5339&type=0 |
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| author | Seema Amin Hyejin Yoon Dong-Hee Choi Yi-Hao Hsu Jungwoo Lee |
| author_facet | Seema Amin Hyejin Yoon Dong-Hee Choi Yi-Hao Hsu Jungwoo Lee |
| author_sort | Seema Amin |
| collection | DOAJ |
| description | Continuous and balanced bone remodeling is essential for maintaining mechanical integrity, mineral homeostasis, and hematopoiesis. Dysregulated bone metabolism develops pathological conditions, such as osteoporosis and bone metastasis. Functional and analytical recapitulation of bone remodeling in vitro is critical for advancing our understanding of bone mineral metabolism, disease mechanisms, and drug development. However, conventional models fail to replicate the essential complexity of the bone extracellular matrix (ECM) and the dynamic interplay between bone-forming osteoblasts and bone-resorbing osteoclasts. Recently, we developed an osteoid-mimicking demineralized bone paper (DBP) by thin-sectioning demineralized bovine compact bone matrix. DBP supports osteoblastic mineral deposition and the subsequent transition to bone-lining cells. When co-cultured with bone marrow mononuclear cells under biochemical stimulation, osteoblasts shift their regulatory secretion profiles and effectively induce osteoclastogenesis. The semi-transparent nature of DBP, combined with primary osteogenic cells retrieved from DsRed and eGFP reporter mice, enables longitudinal fluorescent monitoring of these multicellular processes and quantitative analysis. In this protocol, we describe the methods for DBP generation, reconstituting mineralized bone tissue complexity with osteoblasts, and recapitulating the bone remodeling cycle through bone marrow monocytes co-culture under biochemical stimulation, offering a useful platform for the related and broader research community. |
| format | Article |
| id | doaj-art-32ef04f6f6a4485d9368c7597f3ec903 |
| institution | Kabale University |
| issn | 2331-8325 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Bio-protocol LLC |
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| series | Bio-Protocol |
| spelling | doaj-art-32ef04f6f6a4485d9368c7597f3ec9032025-08-20T03:26:34ZengBio-protocol LLCBio-Protocol2331-83252025-06-01151110.21769/BioProtoc.5339Murine Osteoblast and Osteoclast Co-culture on Demineralized Bone Paper for Bone RemodelingSeema Amin0Hyejin Yoon1Dong-Hee Choi2Yi-Hao Hsu3Jungwoo Lee4Molecular & Cellular Biology Graduate Program, University of Massachusetts, Amherst, MA, USADepartment of Biomedical Engineering, University of Massachusetts, Amherst, MA, USADepartment of Chemical Engineering, University of Massachusetts, Amherst, MA, USAMolecular & Cellular Biology Graduate Program, University of Massachusetts, Amherst, MA, USAMolecular & Cellular Biology Graduate Program, University of Massachusetts, Amherst, MA, USADepartment of Biomedical Engineering, University of Massachusetts, Amherst, MA, USA, Department of Chemical Engineering, University of Massachusetts, Amherst, MA, USAContinuous and balanced bone remodeling is essential for maintaining mechanical integrity, mineral homeostasis, and hematopoiesis. Dysregulated bone metabolism develops pathological conditions, such as osteoporosis and bone metastasis. Functional and analytical recapitulation of bone remodeling in vitro is critical for advancing our understanding of bone mineral metabolism, disease mechanisms, and drug development. However, conventional models fail to replicate the essential complexity of the bone extracellular matrix (ECM) and the dynamic interplay between bone-forming osteoblasts and bone-resorbing osteoclasts. Recently, we developed an osteoid-mimicking demineralized bone paper (DBP) by thin-sectioning demineralized bovine compact bone matrix. DBP supports osteoblastic mineral deposition and the subsequent transition to bone-lining cells. When co-cultured with bone marrow mononuclear cells under biochemical stimulation, osteoblasts shift their regulatory secretion profiles and effectively induce osteoclastogenesis. The semi-transparent nature of DBP, combined with primary osteogenic cells retrieved from DsRed and eGFP reporter mice, enables longitudinal fluorescent monitoring of these multicellular processes and quantitative analysis. In this protocol, we describe the methods for DBP generation, reconstituting mineralized bone tissue complexity with osteoblasts, and recapitulating the bone remodeling cycle through bone marrow monocytes co-culture under biochemical stimulation, offering a useful platform for the related and broader research community.https://bio-protocol.org/en/bpdetail?id=5339&type=0 |
| spellingShingle | Seema Amin Hyejin Yoon Dong-Hee Choi Yi-Hao Hsu Jungwoo Lee Murine Osteoblast and Osteoclast Co-culture on Demineralized Bone Paper for Bone Remodeling Bio-Protocol |
| title | Murine Osteoblast and Osteoclast Co-culture on Demineralized Bone Paper for Bone Remodeling |
| title_full | Murine Osteoblast and Osteoclast Co-culture on Demineralized Bone Paper for Bone Remodeling |
| title_fullStr | Murine Osteoblast and Osteoclast Co-culture on Demineralized Bone Paper for Bone Remodeling |
| title_full_unstemmed | Murine Osteoblast and Osteoclast Co-culture on Demineralized Bone Paper for Bone Remodeling |
| title_short | Murine Osteoblast and Osteoclast Co-culture on Demineralized Bone Paper for Bone Remodeling |
| title_sort | murine osteoblast and osteoclast co culture on demineralized bone paper for bone remodeling |
| url | https://bio-protocol.org/en/bpdetail?id=5339&type=0 |
| work_keys_str_mv | AT seemaamin murineosteoblastandosteoclastcocultureondemineralizedbonepaperforboneremodeling AT hyejinyoon murineosteoblastandosteoclastcocultureondemineralizedbonepaperforboneremodeling AT dongheechoi murineosteoblastandosteoclastcocultureondemineralizedbonepaperforboneremodeling AT yihaohsu murineosteoblastandosteoclastcocultureondemineralizedbonepaperforboneremodeling AT jungwoolee murineosteoblastandosteoclastcocultureondemineralizedbonepaperforboneremodeling |