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Article: Harmonization and standardization of nucleus pulposus cell extraction and culture methods
Title | Harmonization and standardization of nucleus pulposus cell extraction and culture methods |
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Authors | Basatvat, ShaghayeghBach, Frances CBarcellona, Marcos NBinch, Abbie LBuckley, Conor TBueno, BrianChahine, Nadeen OChee, AnaCreemers, Laura BDudli, StefanFearing, BaileyFerguson, Stephen JGansau, JenniferGantenbein, BenjaminGawri, RahulGlaeser, Juliane DGrad, SibylleGuerrero, JulienHaglund, LisbetHernandez, Paula AHoyland, Judith AHuang, CharlesIatridis, James CIllien-Junger, SvenjaJing, LiufangKraus, PetraLaagland, Lisanne TLang, GernotLeung, VictorLi, ZhenLufkin, ThomasVan Maanen, Josette CMcDonnell, Emily EPanebianco, Chris JPresciutti, Steven MRao, SanjnaRichardson, Stephen MRomereim, SarahSchmitz, Tara CSchol, JordySetton, LoriSheyn, DmitriySnuggs, Joseph WSun, YTan, XiaohongTryfonidou, Marianna AVo, NamWang, DongWilliams, BrandonWilliams, RebeccaYoon, S TimLe Maitre, Christine L |
Keywords | culture harmonization in vitro intervertebral nucleus pulposus standardization |
Issue Date | 10-Jan-2023 |
Publisher | Wiley Open Access |
Citation | JOR Spine, 2023, v. 6, n. 1 How to Cite? |
Abstract | BackgroundIn 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. MethodsThe 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. ResultsRecommended protocols are provided for extraction, expansion, and re-differentiation of NP cells from common species utilized for NP cell culture. ConclusionsThis 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. |
Persistent Identifier | http://hdl.handle.net/10722/338697 |
ISSN | 2023 Impact Factor: 3.4 2023 SCImago Journal Rankings: 0.892 |
ISI Accession Number ID |
DC Field | Value | Language |
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dc.contributor.author | Basatvat, Shaghayegh | - |
dc.contributor.author | Bach, Frances C | - |
dc.contributor.author | Barcellona, Marcos N | - |
dc.contributor.author | Binch, Abbie L | - |
dc.contributor.author | Buckley, Conor T | - |
dc.contributor.author | Bueno, Brian | - |
dc.contributor.author | Chahine, Nadeen O | - |
dc.contributor.author | Chee, Ana | - |
dc.contributor.author | Creemers, Laura B | - |
dc.contributor.author | Dudli, Stefan | - |
dc.contributor.author | Fearing, Bailey | - |
dc.contributor.author | Ferguson, Stephen J | - |
dc.contributor.author | Gansau, Jennifer | - |
dc.contributor.author | Gantenbein, Benjamin | - |
dc.contributor.author | Gawri, Rahul | - |
dc.contributor.author | Glaeser, Juliane D | - |
dc.contributor.author | Grad, Sibylle | - |
dc.contributor.author | Guerrero, Julien | - |
dc.contributor.author | Haglund, Lisbet | - |
dc.contributor.author | Hernandez, Paula A | - |
dc.contributor.author | Hoyland, Judith A | - |
dc.contributor.author | Huang, Charles | - |
dc.contributor.author | Iatridis, James C | - |
dc.contributor.author | Illien-Junger, Svenja | - |
dc.contributor.author | Jing, Liufang | - |
dc.contributor.author | Kraus, Petra | - |
dc.contributor.author | Laagland, Lisanne T | - |
dc.contributor.author | Lang, Gernot | - |
dc.contributor.author | Leung, Victor | - |
dc.contributor.author | Li, Zhen | - |
dc.contributor.author | Lufkin, Thomas | - |
dc.contributor.author | Van Maanen, Josette C | - |
dc.contributor.author | McDonnell, Emily E | - |
dc.contributor.author | Panebianco, Chris J | - |
dc.contributor.author | Presciutti, Steven M | - |
dc.contributor.author | Rao, Sanjna | - |
dc.contributor.author | Richardson, Stephen M | - |
dc.contributor.author | Romereim, Sarah | - |
dc.contributor.author | Schmitz, Tara C | - |
dc.contributor.author | Schol, Jordy | - |
dc.contributor.author | Setton, Lori | - |
dc.contributor.author | Sheyn, Dmitriy | - |
dc.contributor.author | Snuggs, Joseph W | - |
dc.contributor.author | Sun, Y | - |
dc.contributor.author | Tan, Xiaohong | - |
dc.contributor.author | Tryfonidou, Marianna A | - |
dc.contributor.author | Vo, Nam | - |
dc.contributor.author | Wang, Dong | - |
dc.contributor.author | Williams, Brandon | - |
dc.contributor.author | Williams, Rebecca | - |
dc.contributor.author | Yoon, S Tim | - |
dc.contributor.author | Le Maitre, Christine L | - |
dc.date.accessioned | 2024-03-11T10:30:51Z | - |
dc.date.available | 2024-03-11T10:30:51Z | - |
dc.date.issued | 2023-01-10 | - |
dc.identifier.citation | JOR Spine, 2023, v. 6, n. 1 | - |
dc.identifier.issn | 2572-1143 | - |
dc.identifier.uri | http://hdl.handle.net/10722/338697 | - |
dc.description.abstract | <h3>Background</h3><p>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.</p><h3>Methods</h3><p>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.</p><h3>Results</h3><p>Recommended protocols are provided for extraction, expansion, and re-differentiation of NP cells from common species utilized for NP cell culture.</p><h3>Conclusions</h3><p>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.</p> | - |
dc.language | eng | - |
dc.publisher | Wiley Open Access | - |
dc.relation.ispartof | JOR Spine | - |
dc.rights | This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. | - |
dc.subject | culture | - |
dc.subject | harmonization | - |
dc.subject | in vitro | - |
dc.subject | intervertebral | - |
dc.subject | nucleus pulposus | - |
dc.subject | standardization | - |
dc.title | Harmonization and standardization of nucleus pulposus cell extraction and culture methods | - |
dc.type | Article | - |
dc.identifier.doi | 10.1002/jsp2.1238 | - |
dc.identifier.scopus | eid_2-s2.0-85146165555 | - |
dc.identifier.volume | 6 | - |
dc.identifier.issue | 1 | - |
dc.identifier.eissn | 2572-1143 | - |
dc.identifier.isi | WOS:000913647800001 | - |
dc.identifier.issnl | 2572-1143 | - |