Harmonization and standardization of nucleus pulposus cell extraction and culture methods.

Autor: Basatvat S; Biomolecular Sciences Research Centre Sheffield Hallam University Sheffield UK., Bach FC; Department of Clinical Sciences, Faculty of Veterinary Medicine Utrecht University Utrecht The Netherlands., Barcellona MN; Trinity Centre for Biomedical Engineering, Trinity Biomedical Sciences Institute, Trinity College Dublin The University of Dublin Dublin Ireland., Binch AL; Biomolecular Sciences Research Centre Sheffield Hallam University Sheffield UK., Buckley CT; Trinity Centre for Biomedical Engineering, Trinity Biomedical Sciences Institute, Trinity College Dublin The University of Dublin Dublin Ireland., Bueno B; Leni & Peter W. May Department of Orthopaedics Icahn School of Medicine at Mount Sinai New York New York USA., Chahine NO; Departments of Orthopedic Surgery and Biomedical Engineering Columbia University New York New York USA., Chee A; Department of Orthopedic Surgery Rush University Medical Center Chicago Illinois USA., Creemers LB; Department of Orthopedics University Medical Center Utrecht Utrecht The Netherlands., Dudli S; Center for Experimental Rheumatology University of Zurich Zurich Switzerland., Fearing B; Department of Orthopedic Surgery Atrium Health Musculoskeletal Institute Charlotte North Carolina USA., Ferguson SJ; Institute for Biomechanics ETH Zurich Zurich Switzerland., Gansau J; Leni & Peter W. May Department of Orthopaedics Icahn School of Medicine at Mount Sinai New York New York USA., Gantenbein B; Bone & Joint Program, Department for BioMedical Research (DBMR), Medical Faculty University of Bern Bern Switzerland.; Department for Orthopedics and Traumatology, Insel University Hospital University of Bern Bern Switzerland., Gawri R; Division of Orthopaedic Surgery, Department of Surgery McGill University Montreal Canada.; Regenerative Orthopaedics and Innovation Laboratory McGill University Montreal Canada., Glaeser JD; Cedars-Sinai Medical Center Los Angeles California USA., Grad S; AO Research Institute Davos Davos Switzerland., Guerrero J; Bone & Joint Program, Department for BioMedical Research (DBMR), Medical Faculty University of Bern Bern Switzerland.; Center of Dental Medicine, Oral Biotechnology & Bioengineering University of Zurich Zurich Switzerland., Haglund L; Division of Orthopaedic Surgery, Department of Surgery McGill University Montreal Canada., Hernandez PA; Department of Orthopaedic Surgery University of Texas Southwestern Medical Center Dallas Texas USA., Hoyland JA; School of Biological Sciences, Faculty of Biology, Medicine and Health, Manchester Academic Health Sciences Centre The University of Manchester Manchester UK., Huang C; Department of Biomedical Engineering University of Miami Coral Gables Florida USA., Iatridis JC; Leni & Peter W. May Department of Orthopaedics Icahn School of Medicine at Mount Sinai New York New York USA., Illien-Junger S; Department of Orthopaedics Emory University School of Medicine Atlanta GA USA., Jing L; Department of Orthopaedics Emory University School of Medicine Atlanta GA USA.; Department of Biomedical Engineering Washington University in St. Louis St. Louis Missouri USA., Kraus P; Department of Orthopaedics Emory University School of Medicine Atlanta GA USA.; Department of Biology Clarkson University Potsdam New York USA., Laagland LT; Department of Clinical Sciences, Faculty of Veterinary Medicine Utrecht University Utrecht The Netherlands., Lang G; Department of Orthopedics and Trauma Surgery, Medical Center, Faculty of Medicine Albert-Ludwigs-University of Freiburg Freiburg im Breisgau Germany., Leung V; Department of Orthopaedics & Traumatology The University of Hong Kong Hong Kong SAR China., Li Z; AO Research Institute Davos Davos Switzerland., Lufkin T; Department of Biology Clarkson University Potsdam New York USA., van Maanen JC; Department of Clinical Sciences, Faculty of Veterinary Medicine Utrecht University Utrecht The Netherlands., McDonnell EE; Trinity Centre for Biomedical Engineering, Trinity Biomedical Sciences Institute, Trinity College Dublin The University of Dublin Dublin Ireland., Panebianco CJ; Leni & Peter W. May Department of Orthopaedics Icahn School of Medicine at Mount Sinai New York New York USA., Presciutti SM; Department of Orthopaedics Emory University School of Medicine Atlanta GA USA., Rao S; Leni & Peter W. May Department of Orthopaedics Icahn School of Medicine at Mount Sinai New York New York USA., Richardson SM; School of Biological Sciences, Faculty of Biology, Medicine and Health, Manchester Academic Health Sciences Centre The University of Manchester Manchester UK., Romereim S; Department of Orthopedic Surgery Atrium Health Musculoskeletal Institute Charlotte North Carolina USA., Schmitz TC; Orthopaedic Biomechanics, Department of Biomedical Engineering Eindhoven University of Technology Eindhoven The Netherlands., Schol J; Department of Orthopedic Surgery Tokai University School of Medicine Isehara Japan., Setton L; Departments of Biomedical Engineering and Orthopedic Surgery Washington University in St. Louis St. Louis Missouri USA., Sheyn D; Cedars-Sinai Medical Center Los Angeles California USA., Snuggs JW; Biomolecular Sciences Research Centre Sheffield Hallam University Sheffield UK., Sun Y; Department of Orthopaedics & Traumatology The University of Hong Kong Hong Kong SAR China., Tan X; Department of Biomedical Engineering Washington University in St. Louis St. Louis Missouri USA., Tryfonidou MA; Department of Clinical Sciences, Faculty of Veterinary Medicine Utrecht University Utrecht The Netherlands., Vo N; Department of Orthopaedic Surgery University of Pittsburgh Pittsburgh Pennsylvania USA., Wang D; Department of Orthopaedic Surgery University of Pittsburgh Pittsburgh Pennsylvania USA., Williams B; Department of Orthopedic Surgery Rush University Medical Center Chicago Illinois USA., Williams R; Biomolecular Sciences Research Centre Sheffield Hallam University Sheffield UK., Yoon ST; Department of Orthopaedics Emory University School of Medicine Atlanta GA USA., Le Maitre CL; Biomolecular Sciences Research Centre Sheffield Hallam University Sheffield UK.; Department of Oncology and Metabolism University of Sheffield Sheffield South Yorkshire UK.
Jazyk: angličtina
Zdroj: JOR spine [JOR Spine] 2023 Jan 10; Vol. 6 (1), pp. e1238. Date of Electronic Publication: 2023 Jan 10 (Print Publication: 2023).
DOI: 10.1002/jsp2.1238
Abstrakt: Background: In vitro studies using nucleus pulposus (NP) cells are commonly used to investigate disc cell biology and pathogenesis, or to aid in the development of new therapies. However, lab-to-lab variability jeopardizes the much-needed progress in the field. Here, an international group of spine scientists collaborated to standardize extraction and expansion techniques for NP cells to reduce variability, improve comparability between labs and improve utilization of funding and resources.
Methods: The most commonly applied methods for NP cell extraction, expansion, and re-differentiation were identified using a questionnaire to research groups worldwide. NP cell extraction methods from rat, rabbit, pig, dog, cow, and human NP tissue were experimentally assessed. Expansion and re-differentiation media and techniques were also investigated.
Results: Recommended protocols are provided for extraction, expansion, and re-differentiation of NP cells from common species utilized for NP cell culture.
Conclusions: This international, multilab and multispecies study identified cell extraction methods for greater cell yield and fewer gene expression changes by applying species-specific pronase usage, 60-100 U/ml collagenase for shorter durations. Recommendations for NP cell expansion, passage number, and many factors driving successful cell culture in different species are also addressed to support harmonization, rigor, and cross-lab comparisons on NP cells worldwide.
Competing Interests: The authors have no relevant conflicts of interest to declare in relation to this article.
(© 2023 The Authors. JOR Spine published by Wiley Periodicals LLC on behalf of Orthopaedic Research Society.)
Databáze: MEDLINE
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