Basatvat, Shaghayegh; Bach, Frances C; Barcellona, Marcos N; Binch, Abbie L; Buckley, Conor T; Bueno, Brian; Chahine, Nadeen O; Chee, Ana; Creemers, Laura B; Dudli, Stefan; Fearing, Bailey; Ferguson, Stephen J; Gansau, Jennifer; Gantenbein, Benjamin; Gawri, Rahul; Glaeser, Juliane D; Grad, Sibylle; Guerrero, Julien; Haglund, Lisbet; Hernandez, Paula A; ... (2023). Harmonization and standardization of nucleus pulposus cell extraction and culture methods. JOR Spine, 6(1), e1238. Wiley 10.1002/jsp2.1238
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JOR_Spine_-_2023_-_Basatvat_-_Harmonization_and_standardization_of_nucleus_pulposus_cell_extraction_and_culture_methods.pdf - Published Version Available under License Creative Commons: Attribution-Noncommercial (CC-BY-NC). Download (21MB) | Preview |
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.
Item Type: |
Journal Article (Original Article) |
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Division/Institute: |
04 Faculty of Medicine > Department of Orthopaedic, Plastic and Hand Surgery (DOPH) > Clinic of Orthopaedic Surgery |
UniBE Contributor: |
Gantenbein, Benjamin |
Subjects: |
600 Technology > 610 Medicine & health |
ISSN: |
2572-1143 |
Publisher: |
Wiley |
Language: |
English |
Submitter: |
Pubmed Import |
Date Deposited: |
31 Mar 2023 10:43 |
Last Modified: |
02 Apr 2023 02:15 |
Publisher DOI: |
10.1002/jsp2.1238 |
PubMed ID: |
36994456 |
Uncontrolled Keywords: |
culture harmonization in vitro intervertebral nucleus pulposus standardization |
BORIS DOI: |
10.48350/181238 |
URI: |
https://boris.unibe.ch/id/eprint/181238 |
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