Harmonization and standardization of nucleus pulposus cell extraction and culture methods

• Type of Material: Article
• Authors: Shaghayegh Basatvat, Sheffield Hallam University; Frances C Bach, Utrecht University; Marcos N Barcellona, The University of Dublin; Abbie L Binch, Sheffield Hallam University; Conor T Buckley, The University of Dublin; Brian Bueno, Icahn School of Medicine at Mount Sinai; Nadeen O Chahine, Columbia University; Ana Chee, Rush University; Laura B Creemers, University Medical Center Utrecht; Stefan Dudli, University of Zurich; Bailey Fearing, Atrium Health Musculoskeletal Institute; Stepen J Ferguson, Institute for Biomechanics, ETH Zurich; Jennifer Gansau, Cedars‐Sinai Medical Center, Los Angeles; Benjamin Gantenbein, University of Bern; Rahul Gawri, McGill University; Juliane D Glaeser, Cedars‐Sinai Medical Center, Los Angeles; Sibylle Grad, University of Zurich; Julien Guerrero, University of Zurich; Lisbet Haglund, McGill University; Paula A Hernandez, University of Texas Southwestern Medical Center, Dallas; Judith A Hoyland, University of Manchester; Charles Huang, University of Miami; James C Iatridis, Icahn School of Medicine at Mount Sinai, New York; Svenja Illien-Junger, Emory University; Liufang Jing, Emory University; Petra Kraus, Emory University; Lisanne T Laagland, Utrecht University; Gernot Lang, Albert‐Ludwigs‐University of Freiburg; Victor Leung, University of Hong Kong; Zhen Li, AO Research Institute Davos; Thomas Lufkin, Clarkson University; Josette C van Maanen, Utrecht University; Emily E McDonnell, The University of Dublin; Chris J Panebianco, Icahn School of Medicine at Mount Sinai; Steven Presciutti, Emory University; Sanjna Rao, Icahn School of Medicine at Mount Sinai; Stephen M Richardson, The University of Manchester; Sarah Romereim, Atrium Health Musculoskeletal Institute; Tara C Schmitz, Eindhoven University of Technology; Jordy Schol, Tokai University; Lori Setton, Washington University in St. Louis; Dmitriy Sheyn, Cedars‐Sinai Medical Center, Los Angeles; Jospeh W Snuggs, Sheffield Hallam University; Y Sun, University of Hong Kong; Xiaohong Tan, Washington University in St. Louis; Marianna A Tryfonidou, Utrecht University; Nam Vo, University of Pittsburgh; Dong Wang, University of Pittsburgh; Brandon Williams, Rush University; Rebecca Williams, Sheffield Hallam University; Sangwook Yoon, Emory University; Christine L Le Maitre, Sheffield Hallam University
• Language: English
• Date: 2023-01-10
• Publisher: WILEY
• Publication Version: Final Published Version
• Copyright Statement: © 2023 The Authors. JOR Spine published by Wiley Periodicals LLC on behalf of Orthopaedic Research Society.
• License: Creative Commons Attribution-NonCommercial 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1002/jsp2.1238
• Title of Journal or Parent Work: JOR SPINE
• Volume: 6
• Issue: 1
• Start Page: e1238
• End Page: e1238
• Keywords: GROWTH; OXYGEN; Life Sciences & Biomedicine; harmonization; culture; standardization; INTERVERTEBRAL DISC; Orthopedics; Science & Technology; GENE-EXPRESSION; LOW-BACK-PAIN; intervertebral; MECHANISM; in vitro; PHENOTYPE; BIOMATERIALS; nucleus pulposus; CHONDROCYTES; DEGENERATION
• Research Categories: Engineering, Biomedical; Health Sciences, Medicine and Surgery

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.