Harmonization and standardization of nucleus pulposus cell extraction and culture methods
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 s...
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Published in | JOR-spine Vol. 6; no. 1; pp. e1238 - n/a |
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Main Authors | , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Hoboken, USA
John Wiley & Sons, Inc
01.03.2023
Wiley |
Subjects | |
Online Access | Get full text |
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Summary: | 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.
An international group of spine scientists collaborated to standardize extraction and expansion techniques for nucleus pulposus cells to reduce variability, improve comparability between labs, and improve utilization of funding and resources. |
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Bibliography: | Funding information AO Spine North America, ThéCell/Réseau de médecine régénérative de McGill (MRM) and The Montreal General Hospital Foundation Code Life; AOSPine International; Berta‐Ottenstein‐Programme for Advanced Clinician Scientists; Dutch Arthritis Foundation, Grant/Award Number: LLP22; H2020 European Institute of Innovation and Technology, Grant/Award Number: 825925; Irish Research Council for Science, Engineering and Technology, Grant/Award Number: GOIPG/2018/2448; Swiss National Science Foundation Project, Grant/Award Number: #310030E_192674/1; US National Institute of Health, Grant/Award Numbers: F32 AR070579, R01‐AR077678, R01‐AR080096, F31‐AR077385; Science Foundation Ireland Career Development Award, Grant/Award Number: 15/CDA/3476; National Science Foundation, Grant/Award Number: DGE‐2139839; Veterans Affairs Career Development Award, Office of Research and Development Biomedical Laboratory Research & Development, Grant/Award Number: IK2‐BX003845; Research Grant Council of Hong Kong, Grant/Award Number: GRF17121619; AOSpine, Grant/Award Number: AOSEAR2020‐02; NIH/NIAMS, Grant/Award Number: K01AR071512 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Funding information AO Spine North America, ThéCell/Réseau de médecine régénérative de McGill (MRM) and The Montreal General Hospital Foundation Code Life; AOSPine International; Berta‐Ottenstein‐Programme for Advanced Clinician Scientists; Dutch Arthritis Foundation, Grant/Award Number: LLP22; H2020 European Institute of Innovation and Technology, Grant/Award Number: 825925; Irish Research Council for Science, Engineering and Technology, Grant/Award Number: GOIPG/2018/2448; Swiss National Science Foundation Project, Grant/Award Number: #310030E_192674/1; US National Institute of Health, Grant/Award Numbers: F32 AR070579, R01‐AR077678, R01‐AR080096, F31‐AR077385; Science Foundation Ireland Career Development Award, Grant/Award Number: 15/CDA/3476; National Science Foundation, Grant/Award Number: DGE‐2139839; Veterans Affairs Career Development Award, Office of Research and Development Biomedical Laboratory Research & Development, Grant/Award Number: IK2‐BX003845; Research Grant Council of Hong Kong, Grant/Award Number: GRF17121619; AOSpine, Grant/Award Number: AOSEAR2020‐02; NIH/NIAMS, Grant/Award Number: K01AR071512 |
ISSN: | 2572-1143 2572-1143 |
DOI: | 10.1002/jsp2.1238 |