Effect of coculturing canine notochordal, nucleus pulposus and mesenchymal stromal cells for intervertebral disc regeneration
Early degenerative changes in the nucleus pulposus (NP) are observed after the disappearance of notochordal cells (NCs). Thus, it has been suggested that NCs play an important role in maintaining the NP and may have a regenerative potential on other cells of the NP. As the number of resident NP cell...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Arkesteijn, Irene T M [verfasserIn] |
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| Format: |
Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2015 |
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| Rechteinformationen: |
Nutzungsrecht: © Arkesteijn et al.; licensee BioMed Central. 2015 |
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| Schlagwörter: |
Mesenchymal Stromal Cells - pathology Culture Media, Conditioned - pharmacology |
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| Übergeordnetes Werk: |
Enthalten in: Arthritis research & therapy - London : BioMed Central, 2003, 17(2015), 1, Seite 60 |
|---|---|
| Übergeordnetes Werk: |
volume:17 ; year:2015 ; number:1 ; pages:60 |
| Links: |
|---|
| DOI / URN: |
10.1186/s13075-015-0569-6 |
|---|
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| 520 | |a Early degenerative changes in the nucleus pulposus (NP) are observed after the disappearance of notochordal cells (NCs). Thus, it has been suggested that NCs play an important role in maintaining the NP and may have a regenerative potential on other cells of the NP. As the number of resident NP cells (NPCs) decreases in a degenerating disc, mesenchymal stromal (stem) cells (MSCs) may be used for cell supplementation. In this study, using cells of one species, the regenerative potential of canine NCs was assessed in long-term three-dimensional coculture with canine NPCs or MSCs. Canine NCs and canine NPCs or MSCs were cocultured in alginate beads for 28 days under hypoxic and high-osmolarity conditions. Cell viability, cell morphology and DNA content, extracellular matrix production and expression of genes related to NC markers (Brachyury, KRT18) and NP matrix production (ACAN, COL2A1, COL1A1) were assessed after 1, 15 and 28 days of culture. NCs did not completely maintain their phenotype (morphology, matrix production, gene expression) during 28 days of culture. In cocultures of NPCs and NCs, both extracellular matrix content and anabolic gene expression remained unchanged compared with monoculture groups, whereas cocultures of MSCs and NCs showed increased glycosaminoglycan/DNA. However, the deposition of these proteoglycans was observed near the NCs and not the MSCs. Brachyury expression in the MSC and NC coculture group increased in time. The latter two findings indicate a trophic effect of MSCs on NCs rather than vice versa. No regenerative potential of canine NCs on canine NPCs or MSCs was observed in this study. However, significant changes in NC phenotype in long-term culture may have resulted in a suboptimal regenerative potential of these NCs. In this respect, NC-conditioned medium may be better than coculture for future studies of the regenerative potential of NCs. | ||
| 540 | |a Nutzungsrecht: © Arkesteijn et al.; licensee BioMed Central. 2015 | ||
| 650 | 4 | |a Notochord - pathology | |
| 650 | 4 | |a Mesenchymal Stromal Cells - pathology | |
| 650 | 4 | |a Culture Media, Conditioned - pharmacology | |
| 650 | 4 | |a Intervertebral Disc - pathology | |
| 650 | 4 | |a Intervertebral Disc Degeneration - pathology | |
| 700 | 1 | |a Smolders, Lucas A |4 oth | |
| 700 | 1 | |a Spillekom, Sandra |4 oth | |
| 700 | 1 | |a Riemers, Frank M |4 oth | |
| 700 | 1 | |a Potier, Esther |4 oth | |
| 700 | 1 | |a Meij, Björn P |4 oth | |
| 700 | 1 | |a Ito, Keita |4 oth | |
| 700 | 1 | |a Tryfonidou, Marianna A |4 oth | |
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10.1186/s13075-015-0569-6 doi PQ20160617 (DE-627)OLC1961017636 (DE-599)GBVOLC1961017636 (PRQ)c1801-377efcf7fd4247fc4c23fc2c4a7b17f68df865035e7570d137020e29abb339163 (KEY)0427448220150000017000100060effectofcoculturingcaninenotochordalnucleuspulposu DE-627 ger DE-627 rakwb eng 610 DNB Arkesteijn, Irene T M verfasserin aut Effect of coculturing canine notochordal, nucleus pulposus and mesenchymal stromal cells for intervertebral disc regeneration 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier Early degenerative changes in the nucleus pulposus (NP) are observed after the disappearance of notochordal cells (NCs). Thus, it has been suggested that NCs play an important role in maintaining the NP and may have a regenerative potential on other cells of the NP. As the number of resident NP cells (NPCs) decreases in a degenerating disc, mesenchymal stromal (stem) cells (MSCs) may be used for cell supplementation. In this study, using cells of one species, the regenerative potential of canine NCs was assessed in long-term three-dimensional coculture with canine NPCs or MSCs. Canine NCs and canine NPCs or MSCs were cocultured in alginate beads for 28 days under hypoxic and high-osmolarity conditions. Cell viability, cell morphology and DNA content, extracellular matrix production and expression of genes related to NC markers (Brachyury, KRT18) and NP matrix production (ACAN, COL2A1, COL1A1) were assessed after 1, 15 and 28 days of culture. NCs did not completely maintain their phenotype (morphology, matrix production, gene expression) during 28 days of culture. In cocultures of NPCs and NCs, both extracellular matrix content and anabolic gene expression remained unchanged compared with monoculture groups, whereas cocultures of MSCs and NCs showed increased glycosaminoglycan/DNA. However, the deposition of these proteoglycans was observed near the NCs and not the MSCs. Brachyury expression in the MSC and NC coculture group increased in time. The latter two findings indicate a trophic effect of MSCs on NCs rather than vice versa. No regenerative potential of canine NCs on canine NPCs or MSCs was observed in this study. However, significant changes in NC phenotype in long-term culture may have resulted in a suboptimal regenerative potential of these NCs. In this respect, NC-conditioned medium may be better than coculture for future studies of the regenerative potential of NCs. Nutzungsrecht: © Arkesteijn et al.; licensee BioMed Central. 2015 Notochord - pathology Mesenchymal Stromal Cells - pathology Culture Media, Conditioned - pharmacology Intervertebral Disc - pathology Intervertebral Disc Degeneration - pathology Smolders, Lucas A oth Spillekom, Sandra oth Riemers, Frank M oth Potier, Esther oth Meij, Björn P oth Ito, Keita oth Tryfonidou, Marianna A oth Enthalten in Arthritis research & therapy London : BioMed Central, 2003 17(2015), 1, Seite 60 (DE-627)363765530 (DE-600)2107602-9 (DE-576)379407604 1478-6354 nnns volume:17 year:2015 number:1 pages:60 http://dx.doi.org/10.1186/s13075-015-0569-6 Volltext http://www.ncbi.nlm.nih.gov/pubmed/25890127 http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=4396569&tool=pmcentrez&rendertype=abstract GBV_USEFLAG_A SYSFLAG_A GBV_OLC SSG-OLC-NED SSG-OLC-PHA SSG-OLC-DE-84 AR 17 2015 1 60 |
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10.1186/s13075-015-0569-6 doi PQ20160617 (DE-627)OLC1961017636 (DE-599)GBVOLC1961017636 (PRQ)c1801-377efcf7fd4247fc4c23fc2c4a7b17f68df865035e7570d137020e29abb339163 (KEY)0427448220150000017000100060effectofcoculturingcaninenotochordalnucleuspulposu DE-627 ger DE-627 rakwb eng 610 DNB Arkesteijn, Irene T M verfasserin aut Effect of coculturing canine notochordal, nucleus pulposus and mesenchymal stromal cells for intervertebral disc regeneration 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier Early degenerative changes in the nucleus pulposus (NP) are observed after the disappearance of notochordal cells (NCs). Thus, it has been suggested that NCs play an important role in maintaining the NP and may have a regenerative potential on other cells of the NP. As the number of resident NP cells (NPCs) decreases in a degenerating disc, mesenchymal stromal (stem) cells (MSCs) may be used for cell supplementation. In this study, using cells of one species, the regenerative potential of canine NCs was assessed in long-term three-dimensional coculture with canine NPCs or MSCs. Canine NCs and canine NPCs or MSCs were cocultured in alginate beads for 28 days under hypoxic and high-osmolarity conditions. Cell viability, cell morphology and DNA content, extracellular matrix production and expression of genes related to NC markers (Brachyury, KRT18) and NP matrix production (ACAN, COL2A1, COL1A1) were assessed after 1, 15 and 28 days of culture. NCs did not completely maintain their phenotype (morphology, matrix production, gene expression) during 28 days of culture. In cocultures of NPCs and NCs, both extracellular matrix content and anabolic gene expression remained unchanged compared with monoculture groups, whereas cocultures of MSCs and NCs showed increased glycosaminoglycan/DNA. However, the deposition of these proteoglycans was observed near the NCs and not the MSCs. Brachyury expression in the MSC and NC coculture group increased in time. The latter two findings indicate a trophic effect of MSCs on NCs rather than vice versa. No regenerative potential of canine NCs on canine NPCs or MSCs was observed in this study. However, significant changes in NC phenotype in long-term culture may have resulted in a suboptimal regenerative potential of these NCs. In this respect, NC-conditioned medium may be better than coculture for future studies of the regenerative potential of NCs. Nutzungsrecht: © Arkesteijn et al.; licensee BioMed Central. 2015 Notochord - pathology Mesenchymal Stromal Cells - pathology Culture Media, Conditioned - pharmacology Intervertebral Disc - pathology Intervertebral Disc Degeneration - pathology Smolders, Lucas A oth Spillekom, Sandra oth Riemers, Frank M oth Potier, Esther oth Meij, Björn P oth Ito, Keita oth Tryfonidou, Marianna A oth Enthalten in Arthritis research & therapy London : BioMed Central, 2003 17(2015), 1, Seite 60 (DE-627)363765530 (DE-600)2107602-9 (DE-576)379407604 1478-6354 nnns volume:17 year:2015 number:1 pages:60 http://dx.doi.org/10.1186/s13075-015-0569-6 Volltext http://www.ncbi.nlm.nih.gov/pubmed/25890127 http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=4396569&tool=pmcentrez&rendertype=abstract GBV_USEFLAG_A SYSFLAG_A GBV_OLC SSG-OLC-NED SSG-OLC-PHA SSG-OLC-DE-84 AR 17 2015 1 60 |
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10.1186/s13075-015-0569-6 doi PQ20160617 (DE-627)OLC1961017636 (DE-599)GBVOLC1961017636 (PRQ)c1801-377efcf7fd4247fc4c23fc2c4a7b17f68df865035e7570d137020e29abb339163 (KEY)0427448220150000017000100060effectofcoculturingcaninenotochordalnucleuspulposu DE-627 ger DE-627 rakwb eng 610 DNB Arkesteijn, Irene T M verfasserin aut Effect of coculturing canine notochordal, nucleus pulposus and mesenchymal stromal cells for intervertebral disc regeneration 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier Early degenerative changes in the nucleus pulposus (NP) are observed after the disappearance of notochordal cells (NCs). Thus, it has been suggested that NCs play an important role in maintaining the NP and may have a regenerative potential on other cells of the NP. As the number of resident NP cells (NPCs) decreases in a degenerating disc, mesenchymal stromal (stem) cells (MSCs) may be used for cell supplementation. In this study, using cells of one species, the regenerative potential of canine NCs was assessed in long-term three-dimensional coculture with canine NPCs or MSCs. Canine NCs and canine NPCs or MSCs were cocultured in alginate beads for 28 days under hypoxic and high-osmolarity conditions. Cell viability, cell morphology and DNA content, extracellular matrix production and expression of genes related to NC markers (Brachyury, KRT18) and NP matrix production (ACAN, COL2A1, COL1A1) were assessed after 1, 15 and 28 days of culture. NCs did not completely maintain their phenotype (morphology, matrix production, gene expression) during 28 days of culture. In cocultures of NPCs and NCs, both extracellular matrix content and anabolic gene expression remained unchanged compared with monoculture groups, whereas cocultures of MSCs and NCs showed increased glycosaminoglycan/DNA. However, the deposition of these proteoglycans was observed near the NCs and not the MSCs. Brachyury expression in the MSC and NC coculture group increased in time. The latter two findings indicate a trophic effect of MSCs on NCs rather than vice versa. No regenerative potential of canine NCs on canine NPCs or MSCs was observed in this study. However, significant changes in NC phenotype in long-term culture may have resulted in a suboptimal regenerative potential of these NCs. In this respect, NC-conditioned medium may be better than coculture for future studies of the regenerative potential of NCs. Nutzungsrecht: © Arkesteijn et al.; licensee BioMed Central. 2015 Notochord - pathology Mesenchymal Stromal Cells - pathology Culture Media, Conditioned - pharmacology Intervertebral Disc - pathology Intervertebral Disc Degeneration - pathology Smolders, Lucas A oth Spillekom, Sandra oth Riemers, Frank M oth Potier, Esther oth Meij, Björn P oth Ito, Keita oth Tryfonidou, Marianna A oth Enthalten in Arthritis research & therapy London : BioMed Central, 2003 17(2015), 1, Seite 60 (DE-627)363765530 (DE-600)2107602-9 (DE-576)379407604 1478-6354 nnns volume:17 year:2015 number:1 pages:60 http://dx.doi.org/10.1186/s13075-015-0569-6 Volltext http://www.ncbi.nlm.nih.gov/pubmed/25890127 http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=4396569&tool=pmcentrez&rendertype=abstract GBV_USEFLAG_A SYSFLAG_A GBV_OLC SSG-OLC-NED SSG-OLC-PHA SSG-OLC-DE-84 AR 17 2015 1 60 |
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10.1186/s13075-015-0569-6 doi PQ20160617 (DE-627)OLC1961017636 (DE-599)GBVOLC1961017636 (PRQ)c1801-377efcf7fd4247fc4c23fc2c4a7b17f68df865035e7570d137020e29abb339163 (KEY)0427448220150000017000100060effectofcoculturingcaninenotochordalnucleuspulposu DE-627 ger DE-627 rakwb eng 610 DNB Arkesteijn, Irene T M verfasserin aut Effect of coculturing canine notochordal, nucleus pulposus and mesenchymal stromal cells for intervertebral disc regeneration 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier Early degenerative changes in the nucleus pulposus (NP) are observed after the disappearance of notochordal cells (NCs). Thus, it has been suggested that NCs play an important role in maintaining the NP and may have a regenerative potential on other cells of the NP. As the number of resident NP cells (NPCs) decreases in a degenerating disc, mesenchymal stromal (stem) cells (MSCs) may be used for cell supplementation. In this study, using cells of one species, the regenerative potential of canine NCs was assessed in long-term three-dimensional coculture with canine NPCs or MSCs. Canine NCs and canine NPCs or MSCs were cocultured in alginate beads for 28 days under hypoxic and high-osmolarity conditions. Cell viability, cell morphology and DNA content, extracellular matrix production and expression of genes related to NC markers (Brachyury, KRT18) and NP matrix production (ACAN, COL2A1, COL1A1) were assessed after 1, 15 and 28 days of culture. NCs did not completely maintain their phenotype (morphology, matrix production, gene expression) during 28 days of culture. In cocultures of NPCs and NCs, both extracellular matrix content and anabolic gene expression remained unchanged compared with monoculture groups, whereas cocultures of MSCs and NCs showed increased glycosaminoglycan/DNA. However, the deposition of these proteoglycans was observed near the NCs and not the MSCs. Brachyury expression in the MSC and NC coculture group increased in time. The latter two findings indicate a trophic effect of MSCs on NCs rather than vice versa. No regenerative potential of canine NCs on canine NPCs or MSCs was observed in this study. However, significant changes in NC phenotype in long-term culture may have resulted in a suboptimal regenerative potential of these NCs. In this respect, NC-conditioned medium may be better than coculture for future studies of the regenerative potential of NCs. Nutzungsrecht: © Arkesteijn et al.; licensee BioMed Central. 2015 Notochord - pathology Mesenchymal Stromal Cells - pathology Culture Media, Conditioned - pharmacology Intervertebral Disc - pathology Intervertebral Disc Degeneration - pathology Smolders, Lucas A oth Spillekom, Sandra oth Riemers, Frank M oth Potier, Esther oth Meij, Björn P oth Ito, Keita oth Tryfonidou, Marianna A oth Enthalten in Arthritis research & therapy London : BioMed Central, 2003 17(2015), 1, Seite 60 (DE-627)363765530 (DE-600)2107602-9 (DE-576)379407604 1478-6354 nnns volume:17 year:2015 number:1 pages:60 http://dx.doi.org/10.1186/s13075-015-0569-6 Volltext http://www.ncbi.nlm.nih.gov/pubmed/25890127 http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=4396569&tool=pmcentrez&rendertype=abstract GBV_USEFLAG_A SYSFLAG_A GBV_OLC SSG-OLC-NED SSG-OLC-PHA SSG-OLC-DE-84 AR 17 2015 1 60 |
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10.1186/s13075-015-0569-6 doi PQ20160617 (DE-627)OLC1961017636 (DE-599)GBVOLC1961017636 (PRQ)c1801-377efcf7fd4247fc4c23fc2c4a7b17f68df865035e7570d137020e29abb339163 (KEY)0427448220150000017000100060effectofcoculturingcaninenotochordalnucleuspulposu DE-627 ger DE-627 rakwb eng 610 DNB Arkesteijn, Irene T M verfasserin aut Effect of coculturing canine notochordal, nucleus pulposus and mesenchymal stromal cells for intervertebral disc regeneration 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier Early degenerative changes in the nucleus pulposus (NP) are observed after the disappearance of notochordal cells (NCs). Thus, it has been suggested that NCs play an important role in maintaining the NP and may have a regenerative potential on other cells of the NP. As the number of resident NP cells (NPCs) decreases in a degenerating disc, mesenchymal stromal (stem) cells (MSCs) may be used for cell supplementation. In this study, using cells of one species, the regenerative potential of canine NCs was assessed in long-term three-dimensional coculture with canine NPCs or MSCs. Canine NCs and canine NPCs or MSCs were cocultured in alginate beads for 28 days under hypoxic and high-osmolarity conditions. Cell viability, cell morphology and DNA content, extracellular matrix production and expression of genes related to NC markers (Brachyury, KRT18) and NP matrix production (ACAN, COL2A1, COL1A1) were assessed after 1, 15 and 28 days of culture. NCs did not completely maintain their phenotype (morphology, matrix production, gene expression) during 28 days of culture. In cocultures of NPCs and NCs, both extracellular matrix content and anabolic gene expression remained unchanged compared with monoculture groups, whereas cocultures of MSCs and NCs showed increased glycosaminoglycan/DNA. However, the deposition of these proteoglycans was observed near the NCs and not the MSCs. Brachyury expression in the MSC and NC coculture group increased in time. The latter two findings indicate a trophic effect of MSCs on NCs rather than vice versa. No regenerative potential of canine NCs on canine NPCs or MSCs was observed in this study. However, significant changes in NC phenotype in long-term culture may have resulted in a suboptimal regenerative potential of these NCs. In this respect, NC-conditioned medium may be better than coculture for future studies of the regenerative potential of NCs. Nutzungsrecht: © Arkesteijn et al.; licensee BioMed Central. 2015 Notochord - pathology Mesenchymal Stromal Cells - pathology Culture Media, Conditioned - pharmacology Intervertebral Disc - pathology Intervertebral Disc Degeneration - pathology Smolders, Lucas A oth Spillekom, Sandra oth Riemers, Frank M oth Potier, Esther oth Meij, Björn P oth Ito, Keita oth Tryfonidou, Marianna A oth Enthalten in Arthritis research & therapy London : BioMed Central, 2003 17(2015), 1, Seite 60 (DE-627)363765530 (DE-600)2107602-9 (DE-576)379407604 1478-6354 nnns volume:17 year:2015 number:1 pages:60 http://dx.doi.org/10.1186/s13075-015-0569-6 Volltext http://www.ncbi.nlm.nih.gov/pubmed/25890127 http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=4396569&tool=pmcentrez&rendertype=abstract GBV_USEFLAG_A SYSFLAG_A GBV_OLC SSG-OLC-NED SSG-OLC-PHA SSG-OLC-DE-84 AR 17 2015 1 60 |
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effect of coculturing canine notochordal, nucleus pulposus and mesenchymal stromal cells for intervertebral disc regeneration |
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Effect of coculturing canine notochordal, nucleus pulposus and mesenchymal stromal cells for intervertebral disc regeneration |
| abstract |
Early degenerative changes in the nucleus pulposus (NP) are observed after the disappearance of notochordal cells (NCs). Thus, it has been suggested that NCs play an important role in maintaining the NP and may have a regenerative potential on other cells of the NP. As the number of resident NP cells (NPCs) decreases in a degenerating disc, mesenchymal stromal (stem) cells (MSCs) may be used for cell supplementation. In this study, using cells of one species, the regenerative potential of canine NCs was assessed in long-term three-dimensional coculture with canine NPCs or MSCs. Canine NCs and canine NPCs or MSCs were cocultured in alginate beads for 28 days under hypoxic and high-osmolarity conditions. Cell viability, cell morphology and DNA content, extracellular matrix production and expression of genes related to NC markers (Brachyury, KRT18) and NP matrix production (ACAN, COL2A1, COL1A1) were assessed after 1, 15 and 28 days of culture. NCs did not completely maintain their phenotype (morphology, matrix production, gene expression) during 28 days of culture. In cocultures of NPCs and NCs, both extracellular matrix content and anabolic gene expression remained unchanged compared with monoculture groups, whereas cocultures of MSCs and NCs showed increased glycosaminoglycan/DNA. However, the deposition of these proteoglycans was observed near the NCs and not the MSCs. Brachyury expression in the MSC and NC coculture group increased in time. The latter two findings indicate a trophic effect of MSCs on NCs rather than vice versa. No regenerative potential of canine NCs on canine NPCs or MSCs was observed in this study. However, significant changes in NC phenotype in long-term culture may have resulted in a suboptimal regenerative potential of these NCs. In this respect, NC-conditioned medium may be better than coculture for future studies of the regenerative potential of NCs. |
| abstractGer |
Early degenerative changes in the nucleus pulposus (NP) are observed after the disappearance of notochordal cells (NCs). Thus, it has been suggested that NCs play an important role in maintaining the NP and may have a regenerative potential on other cells of the NP. As the number of resident NP cells (NPCs) decreases in a degenerating disc, mesenchymal stromal (stem) cells (MSCs) may be used for cell supplementation. In this study, using cells of one species, the regenerative potential of canine NCs was assessed in long-term three-dimensional coculture with canine NPCs or MSCs. Canine NCs and canine NPCs or MSCs were cocultured in alginate beads for 28 days under hypoxic and high-osmolarity conditions. Cell viability, cell morphology and DNA content, extracellular matrix production and expression of genes related to NC markers (Brachyury, KRT18) and NP matrix production (ACAN, COL2A1, COL1A1) were assessed after 1, 15 and 28 days of culture. NCs did not completely maintain their phenotype (morphology, matrix production, gene expression) during 28 days of culture. In cocultures of NPCs and NCs, both extracellular matrix content and anabolic gene expression remained unchanged compared with monoculture groups, whereas cocultures of MSCs and NCs showed increased glycosaminoglycan/DNA. However, the deposition of these proteoglycans was observed near the NCs and not the MSCs. Brachyury expression in the MSC and NC coculture group increased in time. The latter two findings indicate a trophic effect of MSCs on NCs rather than vice versa. No regenerative potential of canine NCs on canine NPCs or MSCs was observed in this study. However, significant changes in NC phenotype in long-term culture may have resulted in a suboptimal regenerative potential of these NCs. In this respect, NC-conditioned medium may be better than coculture for future studies of the regenerative potential of NCs. |
| abstract_unstemmed |
Early degenerative changes in the nucleus pulposus (NP) are observed after the disappearance of notochordal cells (NCs). Thus, it has been suggested that NCs play an important role in maintaining the NP and may have a regenerative potential on other cells of the NP. As the number of resident NP cells (NPCs) decreases in a degenerating disc, mesenchymal stromal (stem) cells (MSCs) may be used for cell supplementation. In this study, using cells of one species, the regenerative potential of canine NCs was assessed in long-term three-dimensional coculture with canine NPCs or MSCs. Canine NCs and canine NPCs or MSCs were cocultured in alginate beads for 28 days under hypoxic and high-osmolarity conditions. Cell viability, cell morphology and DNA content, extracellular matrix production and expression of genes related to NC markers (Brachyury, KRT18) and NP matrix production (ACAN, COL2A1, COL1A1) were assessed after 1, 15 and 28 days of culture. NCs did not completely maintain their phenotype (morphology, matrix production, gene expression) during 28 days of culture. In cocultures of NPCs and NCs, both extracellular matrix content and anabolic gene expression remained unchanged compared with monoculture groups, whereas cocultures of MSCs and NCs showed increased glycosaminoglycan/DNA. However, the deposition of these proteoglycans was observed near the NCs and not the MSCs. Brachyury expression in the MSC and NC coculture group increased in time. The latter two findings indicate a trophic effect of MSCs on NCs rather than vice versa. No regenerative potential of canine NCs on canine NPCs or MSCs was observed in this study. However, significant changes in NC phenotype in long-term culture may have resulted in a suboptimal regenerative potential of these NCs. In this respect, NC-conditioned medium may be better than coculture for future studies of the regenerative potential of NCs. |
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Effect of coculturing canine notochordal, nucleus pulposus and mesenchymal stromal cells for intervertebral disc regeneration |
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Smolders, Lucas A Spillekom, Sandra Riemers, Frank M Potier, Esther Meij, Björn P Ito, Keita Tryfonidou, Marianna A |
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