Mature primary human osteocytes in mini organotypic cultures secrete FGF23 and PTH1-34-regulated sclerostin

Frontiers in Endocrinology

Endocrinology

Helen J. Knowles¹  Philippa A. Hulley¹  Anastasios Chanalaris¹  Adam P. Cribbs²  Liam M. Grover³  Argyro Koutsikouni⁴ 

[1] Botnar Institute for Musculoskeletal Sciences, Nuffield Department of Orthopaedics Rheumatology and Musculoskeletal Sciences, University of Oxford, Oxford, United Kingdom;Botnar Institute for Musculoskeletal Sciences, Nuffield Department of Orthopaedics Rheumatology and Musculoskeletal Sciences, University of Oxford, Oxford, United Kingdom;Oxford Centre for Translational Myeloma Research, Botnar Institute for Musculoskeletal Sciences, University of Oxford, Oxford, United Kingdom;Healthcare Technologies Institute, School of Chemical Engineering, University of Birmingham, Birmingham, United Kingdom;School of Biology, Aristotle University of Thessaloniki, Thessaloniki, Greece;

关键词: osteocyte;    sclerostin;    FGF23;    mineralization;    osteoclast;    organoid;   

DOI  :  10.3389/fendo.2023.1167734

 received in 2023-02-16, accepted in 2023-04-03,  发布年份 2023

来源: Frontiers

【 摘 要 】

IntroductionFor decades, functional primary human osteocyte cultures have been crucially needed for understanding their role in bone anabolic processes and in endocrine phosphate regulation via the bone-kidney axis. Mature osteocyte proteins (sclerostin, DMP1, Phex and FGF23) play a key role in various systemic diseases and are targeted by successful bone anabolic drugs (anti-sclerostin antibody and teriparatide (PTH1-34)). However, cell lines available to study osteocytes produce very little sclerostin and low levels of mature osteocyte markers. We have developed a primary human 3D organotypic culture system that replicates the formation of mature osteocytes in bone.MethodsPrimary human osteoblasts were seeded in a fibrinogen / thrombin gel around 3D-printed hanging posts. Following contraction of the gel around the posts, cells were cultured in osteogenic media and conditioned media was collected for analysis of secreted markers of osteocyte formation.ResultsThe organoids were viable for at least 6 months, allowing co-culture with different cell types and testing of bone anabolic drugs. Bulk RNAseq data displayed the developing marker trajectory of ossification and human primary osteocyte formation in vitro over an initial 8- week period. Vitamin D3 supplementation increased mineralization and sclerostin secretion, while hypoxia and PTH1-34 modulated sclerostin. Our culture system also secreted FGF23, enabling the future development of a bone-kidney-parathyroid-vascular multi-organoid or organ-on-a-chip system to study disease processes and drug effects using purely human cells.DiscussionThis 3D organotypic culture system provides a stable, long-lived, and regulated population of mature human primary osteocytes for a variety of research applications.

【 授权许可】

Unknown   
Copyright © 2023 Knowles, Chanalaris, Koutsikouni, Cribbs, Grover and Hulley