Intervertebral disc and endplate cell characterisation highlights annulus fibrosus cells as the most promising for tissue-specific disc degeneration therapy: European Cells and Materials

P. De Luca; M. Castagnetta; L. de Girolamo; S. Coco; M. Malacarne; E. Ragni; M. Viganò; G. Lugano; M. Brayda-Bruno; D. Coviello; A. Colombini

doi:10.22203/eCM.v039a10

Intervertebral disc and endplate cell characterisation highlights annulus fibrosus cells as the most promising for tissue-specific disc degeneration therapy: European Cells and Materials

P. De Luca, M. Castagnetta, L. de Girolamo, S. Coco, M. Malacarne, E. Ragni, M. Viganò, G. Lugano, M. Brayda-Bruno, D. Coviello, A. Colombini

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

Abstract

Degenerative processes of the intervertebral disc (IVD) and cartilaginous endplate lead to chronic spine pathologies. Several studies speculated on the intrinsic regenerative capacity of degenerated IVD related to the presence of local mesenchymal progenitors. However, a complete characterisation of the resident IVD cell populations, particularly that isolated from the endplate, is lacking. The purpose of the present study was to characterise the gene expression profiles of human nucleus pulposus (NPCs), annulus fibrosus (AFCs) and endplate (EPCs) cells, setting the basis for future studies aimed at identifying the most promising cells for regenerative purposes. Cells isolated from NP, AF and EP were analysed after vo expansion for their stemness ability, immunophenotype and gene profiles by large-scale microarray analysis. The three cell populations shared a similar clonogenic, adipogenic and osteogenic potential, as well as an immunophenotype with a pattern resembling that of mesenchymal stem cells. NPCs maintained the greatest chondrogenic potential and shared with EPCs the loss of proliferation capability during expansion. The largest number of selectively highly expressed stemness, chondrogenic/tissue-specific and surface genes was found in AFCs, thus representing the most promising source of tissue-specific expanded cells for the treatment of IVD degeneration. © 2020, AO Research Institute Davos. All rights reserved.

Original language	English
Pages (from-to)	156-170
Number of pages	15
Journal	Eur. Cells and Mater.
Volume	39
DOIs	https://doi.org/10.22203/eCM.v039a10
Publication status	Published - 2020

Keywords

Annulus fibrosus
Endplate
Intervertebral disc
Microarray
Nucleus pulposus
Phenotype
adipogenesis
adult
annulus fibrosus
Article
bone development
cell aging
cell differentiation
cell expansion
cell function
cell isolation
cell population
cell proliferation
cell therapy
clinical article
clonogenesis
controlled study
female
gene expression profiling
human
human cell
human tissue
immunophenotyping
in vitro study
intervertebral disk degeneration
male
microarray analysis
middle aged
nerve ending
nucleus pulposus
RNA isolation
surgical technique

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10.22203/eCM.v039a10

https://www.scopus.com/inward/record.uri?eid=2-s2.0-85081089141&doi=10.22203%2feCM.v039a10&partnerID=40&md5=bfcf2e6c19816d25513098589d78f13b

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De Luca, P., Castagnetta, M., de Girolamo, L., Coco, S., Malacarne, M., Ragni, E., Viganò, M., Lugano, G., Brayda-Bruno, M., Coviello, D., & Colombini, A. (2020). Intervertebral disc and endplate cell characterisation highlights annulus fibrosus cells as the most promising for tissue-specific disc degeneration therapy: European Cells and Materials. Eur. Cells and Mater., 39, 156-170. https://doi.org/10.22203/eCM.v039a10

Intervertebral disc and endplate cell characterisation highlights annulus fibrosus cells as the most promising for tissue-specific disc degeneration therapy : European Cells and Materials. / De Luca, P.; Castagnetta, M.; de Girolamo, L.; Coco, S.; Malacarne, M.; Ragni, E.; Viganò, M.; Lugano, G.; Brayda-Bruno, M.; Coviello, D.; Colombini, A.

In: Eur. Cells and Mater., Vol. 39, 2020, p. 156-170.

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

De Luca, P, Castagnetta, M, de Girolamo, L , Coco, S , Malacarne, M, Ragni, E, Viganò, M, Lugano, G, Brayda-Bruno, M , Coviello, D & Colombini, A 2020, 'Intervertebral disc and endplate cell characterisation highlights annulus fibrosus cells as the most promising for tissue-specific disc degeneration therapy: European Cells and Materials', Eur. Cells and Mater., vol. 39, pp. 156-170. https://doi.org/10.22203/eCM.v039a10

De Luca, P. ; Castagnetta, M. ; de Girolamo, L. ; Coco, S. ; Malacarne, M. ; Ragni, E. ; Viganò, M. ; Lugano, G. ; Brayda-Bruno, M. ; Coviello, D. ; Colombini, A. / Intervertebral disc and endplate cell characterisation highlights annulus fibrosus cells as the most promising for tissue-specific disc degeneration therapy : European Cells and Materials. In: Eur. Cells and Mater. 2020 ; Vol. 39. pp. 156-170.

@article{1be31ef013ba4bd8acd995e565d8af26,

title = "Intervertebral disc and endplate cell characterisation highlights annulus fibrosus cells as the most promising for tissue-specific disc degeneration therapy: European Cells and Materials",

abstract = "Degenerative processes of the intervertebral disc (IVD) and cartilaginous endplate lead to chronic spine pathologies. Several studies speculated on the intrinsic regenerative capacity of degenerated IVD related to the presence of local mesenchymal progenitors. However, a complete characterisation of the resident IVD cell populations, particularly that isolated from the endplate, is lacking. The purpose of the present study was to characterise the gene expression profiles of human nucleus pulposus (NPCs), annulus fibrosus (AFCs) and endplate (EPCs) cells, setting the basis for future studies aimed at identifying the most promising cells for regenerative purposes. Cells isolated from NP, AF and EP were analysed after vo expansion for their stemness ability, immunophenotype and gene profiles by large-scale microarray analysis. The three cell populations shared a similar clonogenic, adipogenic and osteogenic potential, as well as an immunophenotype with a pattern resembling that of mesenchymal stem cells. NPCs maintained the greatest chondrogenic potential and shared with EPCs the loss of proliferation capability during expansion. The largest number of selectively highly expressed stemness, chondrogenic/tissue-specific and surface genes was found in AFCs, thus representing the most promising source of tissue-specific expanded cells for the treatment of IVD degeneration. {\textcopyright} 2020, AO Research Institute Davos. All rights reserved.",

keywords = "Annulus fibrosus, Endplate, Intervertebral disc, Microarray, Nucleus pulposus, Phenotype, adipogenesis, adult, annulus fibrosus, Article, bone development, cell aging, cell differentiation, cell expansion, cell function, cell isolation, cell population, cell proliferation, cell therapy, clinical article, clonogenesis, controlled study, female, gene expression profiling, human, human cell, human tissue, immunophenotyping, in vitro study, intervertebral disk degeneration, male, microarray analysis, middle aged, nerve ending, nucleus pulposus, RNA isolation, surgical technique",

author = "{De Luca}, P. and M. Castagnetta and {de Girolamo}, L. and S. Coco and M. Malacarne and E. Ragni and M. Vigan{\`o} and G. Lugano and M. Brayda-Bruno and D. Coviello and A. Colombini",

note = "Cited By :2 Export Date: 24 February 2021 Correspondence Address: de Girolamo, L.via Riccardo Galeazzi 4, Italy; email: laura.degirolamo@grupposandonato.it Funding text 1: The study was funded by the Italian Ministry of Funding text 2: Health, “Ricerca Corrente”. S.C. is a PhD supported by grants from the Italian Ministry of Health (Ricerca Corrente). The authors declare no conflict of interest. References: Bendtsen, M., Bunger, C., Colombier, P., Le Visage, C., Roberts, S., Sakai, D., Urban, J.P.G., Biological challenges for regeneration of the degenerated disc using cellular therapies (2016) Acta Orthop, 87, pp. 39-46; Bendtsen, M., Bunger, C.E., Zou, X.N., Foldager, C., Jorgensen, H.S., Autologous stem cell therapy maintains vertebral blood flow and contrast diffusion through the endplate in experimental intervertebral disc degeneration (2011) Spine, 36, pp. E373-E379; Blanco, J.F., Graciani, I.F., Sanchez-Guijo, F.M., Muntion, S., Hernandez-Campo, P., Santamaria, C., Carrancio, S., Del Canizo, M.C., Isolation and characterization of mesenchymal stromal cells from human degenerated nucleus pulposus: Comparison with bone marrow mesenchymal stromal cells from the same subjects (2010) Spine (Phila Pa 1976), 35, pp. 2259-2265; Brisby, H., Papadimitriou, N., Brantsing, C., Bergh, P., Lindahl, A., Barreto Henriksson, H., The presence of local mesenchymal progenitor cells in human degenerated intervertebral discs and possibilities to influence these vo: A descriptive study in humans (2013) Stem Cells Dev, 22, pp. 804-814; Brown, S., Matta, A., Erwin, M., Roberts, S., Gruber, H.E., Hanley, E.N., Jr., Little, C.B., Melrose, J., Cell clusters are indicative of stem cell activity in the degenerate intervertebral disc: Can their properties be manipulated to improve intrinsic repair of the disc? (2018) Stem Cells Dev, 27, pp. 147-165; Colombini, A., Lanteri, P., Lombardi, G., Grasso, D., Recordati, C., Lovi, A., Banfi, G., Brayda-Bruno, M., Metabolic effects of vitamin D active metabolites in monolayer and micromass cultures of nucleus pulposus and annulus fibrosus cells isolated from human intervertebral disc (2012) Int J Biochem Cell Biol, 44, pp. 1019-1030; Colombini, A., Lombardi, G., Corsi, M.M., Banfi, G., Pathophysiology of the human intervertebral disc (2008) Int J Biochem Cell Biol, 40, pp. 837-842; Colombini, A., Lopa, S., Ceriani, C., Lovati, A.B., Croiset, S.J., Di Giancamillo, A., Lombardi, G., Moretti, M., I vo characterization and vvo behavior of human nucleus pulposus and annulus fibrosus cells in clinical-grade fibrin and collagen-enriched fibrin gels (2015) Tissue Eng Part A, 21, pp. 793-802; de Girolamo, L., Lopa, S., Arrigoni, E., Sartori, M.F., Preis, F.W.B., Brini, A.T., Human adipose-derived stem cells isolated from young and elderly women: Their differentiation potential and scaffold interaction during vo osteoblastic differentiation (2009) Cytotherapy, 11, pp. 793-803; de Luca, P., Kouroupis, D., Vigano, M., Perucca-Orfei, C., Kaplan, L., Zagra, L., de Girolamo, L., Colombini, A., Human diseased articular cartilage contains a mesenchymal stem cell-like population of chondroprogenitors with strong immunomodulatory responses (2019) J Clin Med, 8; Dominici, M., Le Blanc, K., Mueller, I., Slaper-Cortenbach, I., Marini, F., Krause, D., Deans, R., Horwitz, E., Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement (2006) Cytotherapy, 8, pp. 315-317; Gruber, H.E., Riley, F.E., Hoelscher, G.L., Ingram, J.A., Bullock, L., Hanley, E.N., Jr., Human annulus progenitor cells: Analyses of this viable endogenous cell population (2016) J Orthop Res, 34, pp. 1351-1360; Huang, B., Liu, L.T., Li, C.Q., Zhuang, Y., Luo, G., Hu, S.Y., Zhou, Y., Study to determine the presence of progenitor cells in the degenerated human cartilage endplates (2012) Eur Spine J, 21, pp. 613-622; Jackson, A.R., Huang, C.Y., Gu, W.Y., Effect of endplate calcification and mechanical deformation on the distribution of glucose in intervertebral disc: A 3D finite element study (2011) Comput Methods Biomech Biomed Engin, 14, pp. 195-204; Keyes, W.M., Mills, A.A., P63: A new link between senescence and aging (2006) Cell Cycle, 5, pp. 260-265; Lee, H.J., Choi, B.H., Min, B.H., Park, S.R., Changes in surface markers of human mesenchymal stem cells during the chondrogenic differentiation and dedifferentiation processes vo (2009) Arthritis Rheum, 60, pp. 2325-2332; Liu, L.T., Huang, B., Li, C.Q., Zhuang, Y., Wang, J., Zhou, Y., Characteristics of stem cells derived from the degenerated human intervertebral disc cartilage endplate (2011) Plos One, 6; Lopa, S., Ceriani, C., Cecchinato, R., Zagra, L., Moretti, M., Colombini, A., Stability of housekeeping genes in human intervertebral disc, endplate and articular cartilage cells in multiple conditions for reliable transcriptional analysis (2016) Eur Cell Mater, 31, pp. 395-406; Lopa, S., Colombini, A., Stanco, D., de Girolamo, L., Sansone, V., Moretti, M., Donor-matched mesenchymal stem cells from knee infrapatellar and subcutaneous adipose tissue of osteoarthritic donors display differential chondrogenic and osteogenic commitment (2014) Eur Cell Mater, 27, pp. 298-311; Lyu, F.J., Cheung, K.M., Zheng, Z., Wang, H., Sakai, D., Leung, V.Y., IVD progenitor cells: A new horizon for understanding disc homeostasis and repair (2019) Nat Rev Rheumatol, 15, pp. 102-112; Nguyen, C., Poiraudeau, S., Rannou, F., Vertebral subchondral bone (2012) Osteoporos Int, 23, pp. S857-S860; Pattappa, G., Li, Z., Peroglio, M., Wismer, N., Alini, M., Grad, S., Diversity of intervertebral disc cells: Phenotype and function (2012) J Anat, 221, pp. 480-496; Pettine, K., Suzuki, R., Sand, T., Murphy, M., Treatment of discogenic back pain with autologous bone marrow concentrate injection with minimum two year follow-up (2016) Int Orthop, 40, pp. 135-140; Pettine, K.A., Murphy, M.B., Suzuki, R.K., Sand, T.T., Percutaneous injection of autologous bone marrow concentrate cells significantly reduces lumbar discogenic pain through 12 months (2015) Stem Cells, 33, pp. 146-156; Pettine, K.A., Suzuki, R.K., Sand, T.T., Murphy, M.B., Autologous bone marrow concentrate intradiscal injection for the treatment of degenerative disc disease with three-year follow-up (2017) Int Orthop, 41, pp. 2097-2103; Power, K.A., Grad, S., Rutges, J.P., Creemers, L.B., van Rijen, M.H., O{\textquoteright}Gaora, P., Wall, J.G., Gallagher, W.M., Identification of cell surface-specific markers to target human nucleus pulposus cells: Expression of carbonic anhydrase XII varies with age and degeneration (2011) Arthritis Rheum, 63, pp. 3876-3886; Rodrigues-Pinto, R., Berry, A., Piper-Hanley, K., Hanley, N., Richardson, S.M., Hoyland, J.A., Spatiotemporal analysis of putative notochordal cell markers reveals CD24 and keratins 8, 18, and 19 as notochord-specific markers during early human intervertebral disc development (2016) J Orthop Res, 34, pp. 1327-1340; Rodrigues-Pinto, R., Ward, L., Humphreys, M., Zeef, L.A.H., Berry, A., Hanley, K.P., Hanley, N., Hoyland, J.A., Human notochordal cell transcriptome unveils potential regulators of cell function in the developing intervertebral disc (2018) Sci Rep, 8; Rutges, J., Creemers, L.B., Dhert, W., Milz, S., Sakai, D., Mochida, J., Alini, M., Grad, S., Variations in gene and protein expression in human nucleus pulposus in comparison to annulus fibrosus and cartilage cells: Potential associations with aging and degeneration (2010) Osteoarthritis Cartilage, 18, pp. 416-423; Sakai, D., Grad, S., Advancing the cellular and molecular therapy for intervertebral disc disease (2015) Adv Drug Deliv Rev, 84, pp. 159-171; Sakai, D., Nakamura, Y., Nakai, T., Mishima, T., Kato, S., Grad, S., Alini, M., Mochida, J., Exhaustion of nucleus pulposus progenitor cells with ageing and degeneration of the intervertebral disc (2012) Nat Commun, 3; Schol, J., Sakai, D., Cell therapy for intervertebral disc herniation and degenerative disc disease: Clinical trials (2019) Int Orthop, 43, pp. 1011-1025; Schroeder, A., Mueller, O., Stocker, S., Salowsky, R., Leiber, M., Gassmann, M., Lightfoot, S., Ragg, T., The RIN: An RNA integrity number for assigning integrity values to RNA measurements (2006) BMC Mol Biol, 7; Schubert, A.K., Smink, J.J., Arp, M., Ringe, J., Hegewald, A.A., Sittinger, M., Quality assessment of surgical disc samples discriminates human annulus fibrosus and nucleus pulposus on tissue and molecular level (2018) Int J Mol Sci, 19; Urban, J.P., Smith, S., Fairbank, J.C., Nutrition of the intervertebral disc (2004) Spine (Phila Pa 1976), 29, pp. 2700-2709; Wang, H., Zhou, Y., Huang, B., Liu, L.T., Liu, M.H., Wang, J., Li, C.Q., Xiong, C.J., Utilization of stem cells in alginate for nucleus pulposus tissue engineering (2014) Tissue Eng Part A, 20, pp. 908-920",

year = "2020",

doi = "10.22203/eCM.v039a10",

language = "English",

volume = "39",

pages = "156--170",

journal = "Eur. Cells and Mater.",

issn = "1473-2262",

publisher = "AO Research Institute Davos",

}

TY - JOUR

T1 - Intervertebral disc and endplate cell characterisation highlights annulus fibrosus cells as the most promising for tissue-specific disc degeneration therapy

T2 - European Cells and Materials

AU - De Luca, P.

AU - Castagnetta, M.

AU - de Girolamo, L.

AU - Coco, S.

AU - Malacarne, M.

AU - Ragni, E.

AU - Viganò, M.

AU - Lugano, G.

AU - Brayda-Bruno, M.

AU - Coviello, D.

AU - Colombini, A.

N1 - Cited By :2 Export Date: 24 February 2021 Correspondence Address: de Girolamo, L.via Riccardo Galeazzi 4, Italy; email: laura.degirolamo@grupposandonato.it Funding text 1: The study was funded by the Italian Ministry of Funding text 2: Health, “Ricerca Corrente”. S.C. is a PhD supported by grants from the Italian Ministry of Health (Ricerca Corrente). The authors declare no conflict of interest. References: Bendtsen, M., Bunger, C., Colombier, P., Le Visage, C., Roberts, S., Sakai, D., Urban, J.P.G., Biological challenges for regeneration of the degenerated disc using cellular therapies (2016) Acta Orthop, 87, pp. 39-46; Bendtsen, M., Bunger, C.E., Zou, X.N., Foldager, C., Jorgensen, H.S., Autologous stem cell therapy maintains vertebral blood flow and contrast diffusion through the endplate in experimental intervertebral disc degeneration (2011) Spine, 36, pp. E373-E379; Blanco, J.F., Graciani, I.F., Sanchez-Guijo, F.M., Muntion, S., Hernandez-Campo, P., Santamaria, C., Carrancio, S., Del Canizo, M.C., Isolation and characterization of mesenchymal stromal cells from human degenerated nucleus pulposus: Comparison with bone marrow mesenchymal stromal cells from the same subjects (2010) Spine (Phila Pa 1976), 35, pp. 2259-2265; Brisby, H., Papadimitriou, N., Brantsing, C., Bergh, P., Lindahl, A., Barreto Henriksson, H., The presence of local mesenchymal progenitor cells in human degenerated intervertebral discs and possibilities to influence these vo: A descriptive study in humans (2013) Stem Cells Dev, 22, pp. 804-814; Brown, S., Matta, A., Erwin, M., Roberts, S., Gruber, H.E., Hanley, E.N., Jr., Little, C.B., Melrose, J., Cell clusters are indicative of stem cell activity in the degenerate intervertebral disc: Can their properties be manipulated to improve intrinsic repair of the disc? (2018) Stem Cells Dev, 27, pp. 147-165; Colombini, A., Lanteri, P., Lombardi, G., Grasso, D., Recordati, C., Lovi, A., Banfi, G., Brayda-Bruno, M., Metabolic effects of vitamin D active metabolites in monolayer and micromass cultures of nucleus pulposus and annulus fibrosus cells isolated from human intervertebral disc (2012) Int J Biochem Cell Biol, 44, pp. 1019-1030; Colombini, A., Lombardi, G., Corsi, M.M., Banfi, G., Pathophysiology of the human intervertebral disc (2008) Int J Biochem Cell Biol, 40, pp. 837-842; Colombini, A., Lopa, S., Ceriani, C., Lovati, A.B., Croiset, S.J., Di Giancamillo, A., Lombardi, G., Moretti, M., I vo characterization and vvo behavior of human nucleus pulposus and annulus fibrosus cells in clinical-grade fibrin and collagen-enriched fibrin gels (2015) Tissue Eng Part A, 21, pp. 793-802; de Girolamo, L., Lopa, S., Arrigoni, E., Sartori, M.F., Preis, F.W.B., Brini, A.T., Human adipose-derived stem cells isolated from young and elderly women: Their differentiation potential and scaffold interaction during vo osteoblastic differentiation (2009) Cytotherapy, 11, pp. 793-803; de Luca, P., Kouroupis, D., Vigano, M., Perucca-Orfei, C., Kaplan, L., Zagra, L., de Girolamo, L., Colombini, A., Human diseased articular cartilage contains a mesenchymal stem cell-like population of chondroprogenitors with strong immunomodulatory responses (2019) J Clin Med, 8; Dominici, M., Le Blanc, K., Mueller, I., Slaper-Cortenbach, I., Marini, F., Krause, D., Deans, R., Horwitz, E., Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement (2006) Cytotherapy, 8, pp. 315-317; Gruber, H.E., Riley, F.E., Hoelscher, G.L., Ingram, J.A., Bullock, L., Hanley, E.N., Jr., Human annulus progenitor cells: Analyses of this viable endogenous cell population (2016) J Orthop Res, 34, pp. 1351-1360; Huang, B., Liu, L.T., Li, C.Q., Zhuang, Y., Luo, G., Hu, S.Y., Zhou, Y., Study to determine the presence of progenitor cells in the degenerated human cartilage endplates (2012) Eur Spine J, 21, pp. 613-622; Jackson, A.R., Huang, C.Y., Gu, W.Y., Effect of endplate calcification and mechanical deformation on the distribution of glucose in intervertebral disc: A 3D finite element study (2011) Comput Methods Biomech Biomed Engin, 14, pp. 195-204; Keyes, W.M., Mills, A.A., P63: A new link between senescence and aging (2006) Cell Cycle, 5, pp. 260-265; Lee, H.J., Choi, B.H., Min, B.H., Park, S.R., Changes in surface markers of human mesenchymal stem cells during the chondrogenic differentiation and dedifferentiation processes vo (2009) Arthritis Rheum, 60, pp. 2325-2332; Liu, L.T., Huang, B., Li, C.Q., Zhuang, Y., Wang, J., Zhou, Y., Characteristics of stem cells derived from the degenerated human intervertebral disc cartilage endplate (2011) Plos One, 6; Lopa, S., Ceriani, C., Cecchinato, R., Zagra, L., Moretti, M., Colombini, A., Stability of housekeeping genes in human intervertebral disc, endplate and articular cartilage cells in multiple conditions for reliable transcriptional analysis (2016) Eur Cell Mater, 31, pp. 395-406; Lopa, S., Colombini, A., Stanco, D., de Girolamo, L., Sansone, V., Moretti, M., Donor-matched mesenchymal stem cells from knee infrapatellar and subcutaneous adipose tissue of osteoarthritic donors display differential chondrogenic and osteogenic commitment (2014) Eur Cell Mater, 27, pp. 298-311; Lyu, F.J., Cheung, K.M., Zheng, Z., Wang, H., Sakai, D., Leung, V.Y., IVD progenitor cells: A new horizon for understanding disc homeostasis and repair (2019) Nat Rev Rheumatol, 15, pp. 102-112; Nguyen, C., Poiraudeau, S., Rannou, F., Vertebral subchondral bone (2012) Osteoporos Int, 23, pp. S857-S860; Pattappa, G., Li, Z., Peroglio, M., Wismer, N., Alini, M., Grad, S., Diversity of intervertebral disc cells: Phenotype and function (2012) J Anat, 221, pp. 480-496; Pettine, K., Suzuki, R., Sand, T., Murphy, M., Treatment of discogenic back pain with autologous bone marrow concentrate injection with minimum two year follow-up (2016) Int Orthop, 40, pp. 135-140; Pettine, K.A., Murphy, M.B., Suzuki, R.K., Sand, T.T., Percutaneous injection of autologous bone marrow concentrate cells significantly reduces lumbar discogenic pain through 12 months (2015) Stem Cells, 33, pp. 146-156; Pettine, K.A., Suzuki, R.K., Sand, T.T., Murphy, M.B., Autologous bone marrow concentrate intradiscal injection for the treatment of degenerative disc disease with three-year follow-up (2017) Int Orthop, 41, pp. 2097-2103; Power, K.A., Grad, S., Rutges, J.P., Creemers, L.B., van Rijen, M.H., O’Gaora, P., Wall, J.G., Gallagher, W.M., Identification of cell surface-specific markers to target human nucleus pulposus cells: Expression of carbonic anhydrase XII varies with age and degeneration (2011) Arthritis Rheum, 63, pp. 3876-3886; Rodrigues-Pinto, R., Berry, A., Piper-Hanley, K., Hanley, N., Richardson, S.M., Hoyland, J.A., Spatiotemporal analysis of putative notochordal cell markers reveals CD24 and keratins 8, 18, and 19 as notochord-specific markers during early human intervertebral disc development (2016) J Orthop Res, 34, pp. 1327-1340; Rodrigues-Pinto, R., Ward, L., Humphreys, M., Zeef, L.A.H., Berry, A., Hanley, K.P., Hanley, N., Hoyland, J.A., Human notochordal cell transcriptome unveils potential regulators of cell function in the developing intervertebral disc (2018) Sci Rep, 8; Rutges, J., Creemers, L.B., Dhert, W., Milz, S., Sakai, D., Mochida, J., Alini, M., Grad, S., Variations in gene and protein expression in human nucleus pulposus in comparison to annulus fibrosus and cartilage cells: Potential associations with aging and degeneration (2010) Osteoarthritis Cartilage, 18, pp. 416-423; Sakai, D., Grad, S., Advancing the cellular and molecular therapy for intervertebral disc disease (2015) Adv Drug Deliv Rev, 84, pp. 159-171; Sakai, D., Nakamura, Y., Nakai, T., Mishima, T., Kato, S., Grad, S., Alini, M., Mochida, J., Exhaustion of nucleus pulposus progenitor cells with ageing and degeneration of the intervertebral disc (2012) Nat Commun, 3; Schol, J., Sakai, D., Cell therapy for intervertebral disc herniation and degenerative disc disease: Clinical trials (2019) Int Orthop, 43, pp. 1011-1025; Schroeder, A., Mueller, O., Stocker, S., Salowsky, R., Leiber, M., Gassmann, M., Lightfoot, S., Ragg, T., The RIN: An RNA integrity number for assigning integrity values to RNA measurements (2006) BMC Mol Biol, 7; Schubert, A.K., Smink, J.J., Arp, M., Ringe, J., Hegewald, A.A., Sittinger, M., Quality assessment of surgical disc samples discriminates human annulus fibrosus and nucleus pulposus on tissue and molecular level (2018) Int J Mol Sci, 19; Urban, J.P., Smith, S., Fairbank, J.C., Nutrition of the intervertebral disc (2004) Spine (Phila Pa 1976), 29, pp. 2700-2709; Wang, H., Zhou, Y., Huang, B., Liu, L.T., Liu, M.H., Wang, J., Li, C.Q., Xiong, C.J., Utilization of stem cells in alginate for nucleus pulposus tissue engineering (2014) Tissue Eng Part A, 20, pp. 908-920

PY - 2020

Y1 - 2020

N2 - Degenerative processes of the intervertebral disc (IVD) and cartilaginous endplate lead to chronic spine pathologies. Several studies speculated on the intrinsic regenerative capacity of degenerated IVD related to the presence of local mesenchymal progenitors. However, a complete characterisation of the resident IVD cell populations, particularly that isolated from the endplate, is lacking. The purpose of the present study was to characterise the gene expression profiles of human nucleus pulposus (NPCs), annulus fibrosus (AFCs) and endplate (EPCs) cells, setting the basis for future studies aimed at identifying the most promising cells for regenerative purposes. Cells isolated from NP, AF and EP were analysed after vo expansion for their stemness ability, immunophenotype and gene profiles by large-scale microarray analysis. The three cell populations shared a similar clonogenic, adipogenic and osteogenic potential, as well as an immunophenotype with a pattern resembling that of mesenchymal stem cells. NPCs maintained the greatest chondrogenic potential and shared with EPCs the loss of proliferation capability during expansion. The largest number of selectively highly expressed stemness, chondrogenic/tissue-specific and surface genes was found in AFCs, thus representing the most promising source of tissue-specific expanded cells for the treatment of IVD degeneration. © 2020, AO Research Institute Davos. All rights reserved.

AB - Degenerative processes of the intervertebral disc (IVD) and cartilaginous endplate lead to chronic spine pathologies. Several studies speculated on the intrinsic regenerative capacity of degenerated IVD related to the presence of local mesenchymal progenitors. However, a complete characterisation of the resident IVD cell populations, particularly that isolated from the endplate, is lacking. The purpose of the present study was to characterise the gene expression profiles of human nucleus pulposus (NPCs), annulus fibrosus (AFCs) and endplate (EPCs) cells, setting the basis for future studies aimed at identifying the most promising cells for regenerative purposes. Cells isolated from NP, AF and EP were analysed after vo expansion for their stemness ability, immunophenotype and gene profiles by large-scale microarray analysis. The three cell populations shared a similar clonogenic, adipogenic and osteogenic potential, as well as an immunophenotype with a pattern resembling that of mesenchymal stem cells. NPCs maintained the greatest chondrogenic potential and shared with EPCs the loss of proliferation capability during expansion. The largest number of selectively highly expressed stemness, chondrogenic/tissue-specific and surface genes was found in AFCs, thus representing the most promising source of tissue-specific expanded cells for the treatment of IVD degeneration. © 2020, AO Research Institute Davos. All rights reserved.

KW - Annulus fibrosus

KW - Endplate

KW - Intervertebral disc

KW - Microarray

KW - Nucleus pulposus

KW - Phenotype

KW - adipogenesis

KW - adult

KW - annulus fibrosus

KW - Article

KW - bone development

KW - cell aging

KW - cell differentiation

KW - cell expansion

KW - cell function

KW - cell isolation

KW - cell population

KW - cell proliferation

KW - cell therapy

KW - clinical article

KW - clonogenesis

KW - controlled study

KW - female

KW - gene expression profiling

KW - human

KW - human cell

KW - human tissue

KW - immunophenotyping

KW - in vitro study

KW - intervertebral disk degeneration

KW - male

KW - microarray analysis

KW - middle aged

KW - nerve ending

KW - nucleus pulposus

KW - RNA isolation

KW - surgical technique

U2 - 10.22203/eCM.v039a10

DO - 10.22203/eCM.v039a10

M3 - Article

VL - 39

SP - 156

EP - 170

JO - Eur. Cells and Mater.

JF - Eur. Cells and Mater.

SN - 1473-2262

ER -

Intervertebral disc and endplate cell characterisation highlights annulus fibrosus cells as the most promising for tissue-specific disc degeneration therapy: European Cells and Materials

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