Harmonization and standardization of nucleus pulposus cell extraction and culture methods.
culture
harmonization
in vitro
intervertebral
nucleus pulposus
standardization
Journal
JOR spine
ISSN: 2572-1143
Titre abrégé: JOR Spine
Pays: United States
ID NLM: 101722350
Informations de publication
Date de publication:
Mar 2023
Mar 2023
Historique:
received:
28
06
2022
revised:
30
10
2022
accepted:
09
12
2022
medline:
31
3
2023
entrez:
30
3
2023
pubmed:
31
3
2023
Statut:
epublish
Résumé
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. 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. Recommended protocols are provided for extraction, expansion, and re-differentiation of NP cells from common species utilized for NP cell culture. 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.
Sections du résumé
Background
UNASSIGNED
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
UNASSIGNED
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
UNASSIGNED
Recommended protocols are provided for extraction, expansion, and re-differentiation of NP cells from common species utilized for NP cell culture.
Conclusions
UNASSIGNED
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.
Identifiants
pubmed: 36994456
doi: 10.1002/jsp2.1238
pii: JSP21238
pmc: PMC10041384
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e1238Subventions
Organisme : NIAMS NIH HHS
ID : K01 AR071512
Pays : United States
Informations de copyright
© 2023 The Authors. JOR Spine published by Wiley Periodicals LLC on behalf of Orthopaedic Research Society.
Déclaration de conflit d'intérêts
The authors have no relevant conflicts of interest to declare in relation to this article.
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