JOR Spine (Mar 2023)
Harmonization and standardization of nucleus pulposus cell extraction and culture methods
- Shaghayegh Basatvat,
- Frances C. Bach,
- Marcos N. Barcellona,
- Abbie L. Binch,
- Conor T. Buckley,
- Brian Bueno,
- Nadeen O. Chahine,
- Ana Chee,
- Laura B. Creemers,
- Stefan Dudli,
- Bailey Fearing,
- Stephen J. Ferguson,
- Jennifer Gansau,
- Benjamin Gantenbein,
- Rahul Gawri,
- Juliane D. Glaeser,
- Sibylle Grad,
- Julien Guerrero,
- Lisbet Haglund,
- Paula A. Hernandez,
- Judith A. Hoyland,
- Charles Huang,
- James C. Iatridis,
- Svenja Illien‐Junger,
- Liufang Jing,
- Petra Kraus,
- Lisanne T. Laagland,
- Gernot Lang,
- Victor Leung,
- Zhen Li,
- Thomas Lufkin,
- Josette C. vanMaanen,
- Emily E. McDonnell,
- Chris J. Panebianco,
- Steven M. Presciutti,
- Sanjna Rao,
- Stephen M. Richardson,
- Sarah Romereim,
- Tara C. Schmitz,
- Jordy Schol,
- Lori Setton,
- Dmitriy Sheyn,
- Joseph W. Snuggs,
- Y. Sun,
- Xiaohong Tan,
- Marianna A. Tryfonidou,
- Nam Vo,
- Dong Wang,
- Brandon Williams,
- Rebecca Williams,
- S. Tim Yoon,
- Christine L. Le Maitre
Affiliations
- Shaghayegh Basatvat
- Biomolecular Sciences Research Centre Sheffield Hallam University Sheffield UK
- Frances C. Bach
- Department of Clinical Sciences, Faculty of Veterinary Medicine Utrecht University Utrecht The Netherlands
- Marcos N. Barcellona
- Trinity Centre for Biomedical Engineering, Trinity Biomedical Sciences Institute, Trinity College Dublin The University of Dublin Dublin Ireland
- Abbie L. Binch
- Biomolecular Sciences Research Centre Sheffield Hallam University Sheffield UK
- Conor T. Buckley
- Trinity Centre for Biomedical Engineering, Trinity Biomedical Sciences Institute, Trinity College Dublin The University of Dublin Dublin Ireland
- Brian Bueno
- Leni & Peter W. May Department of Orthopaedics Icahn School of Medicine at Mount Sinai New York New York USA
- Nadeen O. Chahine
- Departments of Orthopedic Surgery and Biomedical Engineering Columbia University New York New York USA
- Ana Chee
- Department of Orthopedic Surgery Rush University Medical Center Chicago Illinois USA
- Laura B. Creemers
- Department of Orthopedics University Medical Center Utrecht Utrecht The Netherlands
- Stefan Dudli
- Center for Experimental Rheumatology University of Zurich Zurich Switzerland
- Bailey Fearing
- Department of Orthopedic Surgery Atrium Health Musculoskeletal Institute Charlotte North Carolina USA
- Stephen J. Ferguson
- Institute for Biomechanics ETH Zurich Zurich Switzerland
- Jennifer Gansau
- Leni & Peter W. May Department of Orthopaedics Icahn School of Medicine at Mount Sinai New York New York USA
- Benjamin Gantenbein
- Bone & Joint Program, Department for BioMedical Research (DBMR), Medical Faculty University of Bern Bern Switzerland
- Rahul Gawri
- Division of Orthopaedic Surgery, Department of Surgery McGill University Montreal Canada
- Juliane D. Glaeser
- Cedars‐Sinai Medical Center Los Angeles California USA
- Sibylle Grad
- AO Research Institute Davos Davos Switzerland
- Julien Guerrero
- Bone & Joint Program, Department for BioMedical Research (DBMR), Medical Faculty University of Bern Bern Switzerland
- Lisbet Haglund
- Division of Orthopaedic Surgery, Department of Surgery McGill University Montreal Canada
- Paula A. Hernandez
- Department of Orthopaedic Surgery University of Texas Southwestern Medical Center Dallas Texas USA
- Judith A. Hoyland
- School of Biological Sciences, Faculty of Biology, Medicine and Health, Manchester Academic Health Sciences Centre The University of Manchester Manchester UK
- Charles Huang
- Department of Biomedical Engineering University of Miami Coral Gables Florida USA
- James C. Iatridis
- Leni & Peter W. May Department of Orthopaedics Icahn School of Medicine at Mount Sinai New York New York USA
- Svenja Illien‐Junger
- Department of Orthopaedics Emory University School of Medicine Atlanta GA USA
- Liufang Jing
- Department of Orthopaedics Emory University School of Medicine Atlanta GA USA
- Petra Kraus
- Department of Orthopaedics Emory University School of Medicine Atlanta GA USA
- Lisanne T. Laagland
- Department of Clinical Sciences, Faculty of Veterinary Medicine Utrecht University Utrecht The Netherlands
- Gernot Lang
- Department of Orthopedics and Trauma Surgery, Medical Center, Faculty of Medicine Albert‐Ludwigs‐University of Freiburg Freiburg im Breisgau Germany
- Victor Leung
- Department of Orthopaedics & Traumatology The University of Hong Kong Hong Kong SAR China
- Zhen Li
- AO Research Institute Davos Davos Switzerland
- Thomas Lufkin
- Department of Biology Clarkson University Potsdam New York USA
- Josette C. vanMaanen
- Department of Clinical Sciences, Faculty of Veterinary Medicine Utrecht University Utrecht The Netherlands
- Emily E. McDonnell
- Trinity Centre for Biomedical Engineering, Trinity Biomedical Sciences Institute, Trinity College Dublin The University of Dublin Dublin Ireland
- Chris J. Panebianco
- Leni & Peter W. May Department of Orthopaedics Icahn School of Medicine at Mount Sinai New York New York USA
- Steven M. Presciutti
- Department of Orthopaedics Emory University School of Medicine Atlanta GA USA
- Sanjna Rao
- Leni & Peter W. May Department of Orthopaedics Icahn School of Medicine at Mount Sinai New York New York USA
- Stephen M. Richardson
- School of Biological Sciences, Faculty of Biology, Medicine and Health, Manchester Academic Health Sciences Centre The University of Manchester Manchester UK
- Sarah Romereim
- Department of Orthopedic Surgery Atrium Health Musculoskeletal Institute Charlotte North Carolina USA
- Tara C. Schmitz
- Orthopaedic Biomechanics, Department of Biomedical Engineering Eindhoven University of Technology Eindhoven The Netherlands
- Jordy Schol
- Department of Orthopedic Surgery Tokai University School of Medicine Isehara Japan
- Lori Setton
- Departments of Biomedical Engineering and Orthopedic Surgery Washington University in St. Louis St. Louis Missouri USA
- Dmitriy Sheyn
- Cedars‐Sinai Medical Center Los Angeles California USA
- Joseph W. Snuggs
- Biomolecular Sciences Research Centre Sheffield Hallam University Sheffield UK
- Y. Sun
- Department of Orthopaedics & Traumatology The University of Hong Kong Hong Kong SAR China
- Xiaohong Tan
- Department of Biomedical Engineering Washington University in St. Louis St. Louis Missouri USA
- Marianna A. Tryfonidou
- Department of Clinical Sciences, Faculty of Veterinary Medicine Utrecht University Utrecht The Netherlands
- Nam Vo
- Department of Orthopaedic Surgery University of Pittsburgh Pittsburgh Pennsylvania USA
- Dong Wang
- Department of Orthopaedic Surgery University of Pittsburgh Pittsburgh Pennsylvania USA
- Brandon Williams
- Department of Orthopedic Surgery Rush University Medical Center Chicago Illinois USA
- Rebecca Williams
- Biomolecular Sciences Research Centre Sheffield Hallam University Sheffield UK
- S. Tim Yoon
- Department of Orthopaedics Emory University School of Medicine Atlanta GA USA
- Christine L. Le Maitre
- Biomolecular Sciences Research Centre Sheffield Hallam University Sheffield UK
- DOI
- https://doi.org/10.1002/jsp2.1238
- Journal volume & issue
-
Vol. 6,
no. 1
pp. n/a – n/a
Abstract
Abstract 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.
Keywords
