Harmonization and standardization of nucleus pulposus cell extraction and culture methods

Shaghayegh Basatvat; Frances C. Bach; Marcos N. Barcellona; Abbie L. Binch; Conor T. Buckley; Brian Bueno; Nadeen O. Chahine; Ana Chee; Laura B. Creemers; Stefan Dudli; Bailey Fearing; Stephen J. Ferguson; Jennifer Gansau; Benjamin Gantenbein; Rahul Gawri; Juliane D. Glaeser; Sibylle Grad; Julien Guerrero; Lisbet Haglund; Paula A. Hernandez; Judith A. Hoyland; Charles Huang; James C. Iatridis; Svenja Illien-Junger; Liufang Jing; Petra Kraus; Lisanne T. Laagland; Gernot Lang; Victor Leung; Zhen Li; Thomas Lufkin; Josette C. van Maanen; Emily E. McDonnell; Chris J. Panebianco; Steven M. Presciutti; Sanjna Rao; Stephen M. Richardson; Sarah Romereim; Tara C. Schmitz; Jordy Schol; Lori Setton; Dmitriy Sheyn; Joseph W. Snuggs; Y. Sun; Xiaohong Tan; Marianna A. Tryfonidou; Nam Vo; Dong Wang; Brandon Williams; Rebecca Williams; S. Tim Yoon; Christine L. Le Maitre

doi:10.1002/jsp2.1238

Harmonization and standardization of nucleus pulposus cell extraction and culture methods

Shaghayegh Basatvat, Frances C. Bach, Marcos N. Barcellona, Abbie L. Binch, Conor T. Buckley, Brian Bueno, Nadeen O. Chahine, Ana Chee, Laura B. Creemers, Stefan Dudli, Bailey Fearing, Stephen J. Ferguson, Jennifer Gansau, Benjamin Gantenbein, Rahul Gawri, Juliane D. Glaeser, Sibylle Grad, Julien Guerrero, Lisbet Haglund, Paula A. HernandezJudith A. Hoyland, Charles Huang, James C. Iatridis, Svenja Illien-Junger, Liufang Jing, Petra Kraus, Lisanne T. Laagland, Gernot Lang, Victor Leung, Zhen Li, Thomas Lufkin, Josette C. van Maanen, Emily E. McDonnell, Chris J. Panebianco, Steven M. Presciutti, Sanjna Rao, Stephen M. Richardson, Sarah Romereim, Tara C. Schmitz, Jordy Schol, Lori Setton, Dmitriy Sheyn, Joseph W. Snuggs, Y. Sun, Xiaohong Tan, Marianna A. Tryfonidou, Nam Vo, Dong Wang, Brandon Williams, Rebecca Williams, S. Tim Yoon, Christine L. Le Maitre

Research output: Contribution to journal › Article › peer-review

10 Scopus citations

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.

Original language	English
Article number	e1238
Journal	JOR Spine
Volume	6
Issue number	1
DOIs	https://doi.org/10.1002/jsp2.1238
State	Published - Mar 2023

Keywords

culture
harmonization
in vitro
intervertebral
nucleus pulposus
standardization

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10.1002/jsp2.1238

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Basatvat, S., Bach, F. C., Barcellona, M. N., Binch, A. L., Buckley, C. T., Bueno, B., Chahine, N. O., Chee, A., Creemers, L. B., Dudli, S., Fearing, B., Ferguson, S. J., Gansau, J., Gantenbein, B., Gawri, R., Glaeser, J. D., Grad, S., Guerrero, J., Haglund, L., ... Le Maitre, C. L. (2023). Harmonization and standardization of nucleus pulposus cell extraction and culture methods. JOR Spine, 6(1), Article e1238. https://doi.org/10.1002/jsp2.1238

@article{ec77b73240ba49e29c11c40ada032134,

title = "Harmonization and standardization of nucleus pulposus cell extraction and culture methods",

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.",

keywords = "culture, harmonization, in vitro, intervertebral, nucleus pulposus, standardization",

author = "Shaghayegh Basatvat and Bach, {Frances C.} and Barcellona, {Marcos N.} and Binch, {Abbie L.} and Buckley, {Conor T.} and Brian Bueno and Chahine, {Nadeen O.} and Ana Chee and Creemers, {Laura B.} and Stefan Dudli and Bailey Fearing and Ferguson, {Stephen J.} and Jennifer Gansau and Benjamin Gantenbein and Rahul Gawri and Glaeser, {Juliane D.} and Sibylle Grad and Julien Guerrero and Lisbet Haglund and Hernandez, {Paula A.} and Hoyland, {Judith A.} and Charles Huang and Iatridis, {James C.} and Svenja Illien-Junger and Liufang Jing and Petra Kraus and Laagland, {Lisanne T.} and Gernot Lang and Victor Leung and Zhen Li and Thomas Lufkin and {van Maanen}, {Josette C.} and McDonnell, {Emily E.} and Panebianco, {Chris J.} and Presciutti, {Steven M.} and Sanjna Rao and Richardson, {Stephen M.} and Sarah Romereim and Schmitz, {Tara C.} and Jordy Schol and Lori Setton and Dmitriy Sheyn and Snuggs, {Joseph W.} and Y. Sun and Xiaohong Tan and Tryfonidou, {Marianna A.} and Nam Vo and Dong Wang and Brandon Williams and Rebecca Williams and Yoon, {S. Tim} and {Le Maitre}, {Christine L.}",

note = "Publisher Copyright: {\textcopyright} 2023 The Authors. JOR Spine published by Wiley Periodicals LLC on behalf of Orthopaedic Research Society.",

year = "2023",

month = mar,

doi = "10.1002/jsp2.1238",

language = "English",

volume = "6",

journal = "JOR Spine",

issn = "2572-1143",

publisher = "Wiley-Blackwell Publishing Ltd",

number = "1",

}

Basatvat, S, Bach, FC, Barcellona, MN, Binch, AL, Buckley, CT, Bueno, B, Chahine, NO, Chee, A, Creemers, LB, Dudli, S, Fearing, B, Ferguson, SJ, Gansau, J, Gantenbein, B, Gawri, R, Glaeser, JD, Grad, S, Guerrero, J, Haglund, L, Hernandez, PA, Hoyland, JA, Huang, C, Iatridis, JC, Illien-Junger, S, Jing, L, Kraus, P, Laagland, LT, Lang, G, Leung, V, Li, Z, Lufkin, T, van Maanen, JC, McDonnell, EE, Panebianco, CJ, Presciutti, SM, Rao, S, Richardson, SM, Romereim, S, Schmitz, TC, Schol, J, Setton, L, Sheyn, D, Snuggs, JW, Sun, Y, Tan, X, Tryfonidou, MA, Vo, N, Wang, D, Williams, B, Williams, R, Yoon, ST & Le Maitre, CL 2023, 'Harmonization and standardization of nucleus pulposus cell extraction and culture methods', JOR Spine, vol. 6, no. 1, e1238. https://doi.org/10.1002/jsp2.1238

TY - JOUR

T1 - Harmonization and standardization of nucleus pulposus cell extraction and culture methods

AU - Basatvat, Shaghayegh

AU - Bach, Frances C.

AU - Barcellona, Marcos N.

AU - Binch, Abbie L.

AU - Buckley, Conor T.

AU - Bueno, Brian

AU - Chahine, Nadeen O.

AU - Chee, Ana

AU - Creemers, Laura B.

AU - Dudli, Stefan

AU - Fearing, Bailey

AU - Ferguson, Stephen J.

AU - Gansau, Jennifer

AU - Gantenbein, Benjamin

AU - Gawri, Rahul

AU - Glaeser, Juliane D.

AU - Grad, Sibylle

AU - Guerrero, Julien

AU - Haglund, Lisbet

AU - Hernandez, Paula A.

AU - Hoyland, Judith A.

AU - Huang, Charles

AU - Iatridis, James C.

AU - Illien-Junger, Svenja

AU - Jing, Liufang

AU - Kraus, Petra

AU - Laagland, Lisanne T.

AU - Lang, Gernot

AU - Leung, Victor

AU - Li, Zhen

AU - Lufkin, Thomas

AU - van Maanen, Josette C.

AU - McDonnell, Emily E.

AU - Panebianco, Chris J.

AU - Presciutti, Steven M.

AU - Rao, Sanjna

AU - Richardson, Stephen M.

AU - Romereim, Sarah

AU - Schmitz, Tara C.

AU - Schol, Jordy

AU - Setton, Lori

AU - Sheyn, Dmitriy

AU - Snuggs, Joseph W.

AU - Sun, Y.

AU - Tan, Xiaohong

AU - Tryfonidou, Marianna A.

AU - Vo, Nam

AU - Wang, Dong

AU - Williams, Brandon

AU - Williams, Rebecca

AU - Yoon, S. Tim

AU - Le Maitre, Christine L.

PY - 2023/3

Y1 - 2023/3

N2 - 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.

AB - 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.

KW - culture

KW - harmonization

KW - in vitro

KW - intervertebral

KW - nucleus pulposus

KW - standardization

UR - http://www.scopus.com/inward/record.url?scp=85146165555&partnerID=8YFLogxK

U2 - 10.1002/jsp2.1238

DO - 10.1002/jsp2.1238

M3 - Article

AN - SCOPUS:85146165555

SN - 2572-1143

VL - 6

JO - JOR Spine

JF - JOR Spine

IS - 1

M1 - e1238

ER -

Harmonization and standardization of nucleus pulposus cell extraction and culture methods

Abstract

Keywords

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