TY - JOUR
T1 - Recommendations for intervertebral disc notochordal cell investigation
T2 - From isolation to characterization
AU - Williams, Rebecca J
AU - Laagland, Lisanne T
AU - Bach, Frances C
AU - Ward, Lizzy
AU - Chan, Wilson
AU - Tam, Vivian
AU - Medzikovic, Adel
AU - Basatvat, Shaghayegh
AU - Paillat, Lily
AU - Vedrenne, Nicolas
AU - Snuggs, Joseph W
AU - Poramba-Liyanage, Deepani W
AU - Hoyland, Judith A
AU - Chan, Danny
AU - Camus, Anne
AU - Richardson, Stephen M
AU - Tryfonidou, Marianna A
AU - Le Maitre, Christine L
N1 - © 2023 The Authors. JOR Spine published by Wiley Periodicals LLC on behalf of Orthopaedic Research Society.
PY - 2023/9/1
Y1 - 2023/9/1
N2 - BACKGROUND: Lineage-tracing experiments have established that the central region of the mature intervertebral disc, the nucleus pulposus (NP), develops from the embryonic structure called "the notochord". However, changes in the cells derived from the notochord which form the NP (i.e., notochordal cells [NCs]), in terms of their phenotype and functional identity from early developmental stages to skeletal maturation are less understood. These key issues require further investigation to better comprehend the role of NCs in homeostasis and degeneration as well as their potential for regeneration. Progress in utilizing NCs is currently hampered due to poor consistency and lack of consensus methodology for in vitro NC extraction, manipulation, and characterization.METHODS: Here, an international group has come together to provide key recommendations and methodologies for NC isolation within key species, numeration, in vitro manipulation and culture, and characterization.RESULTS: Recommeded protocols are provided for isolation and culture of NCs. Experimental testing provided recommended methodology for numeration of NCs. The issues of cryopreservation are demonstrated, and a pannel of immunohistochemical markers are provided to inform NC characterization.CONCLUSIONS: Together we hope this article provides a road map for in vitro studies of NCs to support advances in research into NC physiology and their potential in regenerative therapies.
AB - BACKGROUND: Lineage-tracing experiments have established that the central region of the mature intervertebral disc, the nucleus pulposus (NP), develops from the embryonic structure called "the notochord". However, changes in the cells derived from the notochord which form the NP (i.e., notochordal cells [NCs]), in terms of their phenotype and functional identity from early developmental stages to skeletal maturation are less understood. These key issues require further investigation to better comprehend the role of NCs in homeostasis and degeneration as well as their potential for regeneration. Progress in utilizing NCs is currently hampered due to poor consistency and lack of consensus methodology for in vitro NC extraction, manipulation, and characterization.METHODS: Here, an international group has come together to provide key recommendations and methodologies for NC isolation within key species, numeration, in vitro manipulation and culture, and characterization.RESULTS: Recommeded protocols are provided for isolation and culture of NCs. Experimental testing provided recommended methodology for numeration of NCs. The issues of cryopreservation are demonstrated, and a pannel of immunohistochemical markers are provided to inform NC characterization.CONCLUSIONS: Together we hope this article provides a road map for in vitro studies of NCs to support advances in research into NC physiology and their potential in regenerative therapies.
KW - culture systems
KW - development
KW - intervertebral disc
KW - notochordal cells
KW - nucleus pulposus
KW - tissue-specific progenitor cells
UR - http://www.scopus.com/inward/record.url?scp=85164597175&partnerID=8YFLogxK
UR - https://www.mendeley.com/catalogue/387b1304-cf4e-3c96-b265-c46457a169be/
U2 - 10.1002/jsp2.1272
DO - 10.1002/jsp2.1272
M3 - Article
C2 - 37780826
SN - 2572-1143
VL - 6
JO - JOR Spine
JF - JOR Spine
IS - 3
M1 - e1272
ER -