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Article: Three-dimensional culture of rabbit nucleus pulposus cells in collagen microspheres

TitleThree-dimensional culture of rabbit nucleus pulposus cells in collagen microspheres
Authors
Keywords3D culture
Collagen
Intervertebral disc
Microencapsulation
Nucleus pulposus cells
Issue Date2011
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/spinee
Citation
Spine Journal, 2011, v. 11 n. 10, p. 947-960 How to Cite?
AbstractBackground: Degenerative disc disease poses an increasing threat to our quality of life as we age. Existing treatments have limitations. New treatment modalities focusing on biologic rather than surgical approach would be appealing. Purpose: Culturing intervertebral disc cells in a three-dimensional (3D) model that can retain cellular characteristics and phenotype is a critical step toward understanding how the disc cells respond to and interact with extrinsic signals before better therapeutics can be derived. Study design: In this work, we studied the culture of rabbit nucleus pulposus (NP) cells in a collagen microsphere system and compared their cell morphology and expression of a few potential phenotypic markers with that in monolayer culture. Methods: Specifically, rabbit NP cells isolated from both young and old animals were encapsulated and cultured in collagen microspheres with different monomeric concentrations and with different cell encapsulation density for different period of time. Evaluation on the growth kinetics, the viability, the cell morphology, the expression of Types I and II collagen, glycosaminoglycans (GAGs), and Keratin 19, and the ultrastructure of the fiber meshwork were conducted to compare the microsphere 3D culture system and the traditional monolayer cultures. Results: Nucleus pulposus cells in two-dimensional culture lost the phenotypic expression of Type II collagen and keratin 19 and expressed Type I collagen. In contrast, the 3D collagen microsphere culture system consistently outperformed the traditional monolayer culture in maintaining a round morphology and preserving the phenotypes of NP cells with persistent expression of Type II collagen and Keratin 19. These cells also remodeled the template collagen matrix in the microspheres by depositing new matrices, including collagen Type II and GAGs in a cell seeding density and collagen concentration dependent manner. Conclusions: This study demonstrates the appeal of the 3D collagen microsphere system for NP cell culture over traditional monolayer culture because it preserves the phenotypic characteristics of NP cells. This system also enables the NP cells to remodel the template collagen matrix by depositing new matrices, suggesting an innovative way to reconstitute cell-specific and native tissue-like environment in vitro for future studies on stem cell matrix niche and interactions of NP cell with extrinsic factors. © 2011 Elsevier Inc. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/139429
ISSN
2015 Impact Factor: 2.66
2015 SCImago Journal Rankings: 1.153
ISI Accession Number ID
Funding AgencyGrant Number
AOSpineAOSBRC-07-06
Research Grant CouncilGRF 760408
Innovation and Technology Commission, the Hong Kong GovernmentITS/552/09FP
University of Hong Kong on Biomedical Engineering
Funding Information:

This work was supported by grants from AOSpine (AOSBRC-07-06); Research Grant Council (GRF 760408); the Innovation and Technology Commission, the Hong Kong Government (ITS/552/09FP) and the Strategic Research Themes of the University of Hong Kong on Biomedical Engineering.

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DC FieldValueLanguage
dc.contributor.authorYuan, Men_HK
dc.contributor.authorLeong, KWen_HK
dc.contributor.authorChan, BPen_HK
dc.date.accessioned2011-09-23T05:49:24Z-
dc.date.available2011-09-23T05:49:24Z-
dc.date.issued2011en_HK
dc.identifier.citationSpine Journal, 2011, v. 11 n. 10, p. 947-960en_HK
dc.identifier.issn1529-9430en_HK
dc.identifier.urihttp://hdl.handle.net/10722/139429-
dc.description.abstractBackground: Degenerative disc disease poses an increasing threat to our quality of life as we age. Existing treatments have limitations. New treatment modalities focusing on biologic rather than surgical approach would be appealing. Purpose: Culturing intervertebral disc cells in a three-dimensional (3D) model that can retain cellular characteristics and phenotype is a critical step toward understanding how the disc cells respond to and interact with extrinsic signals before better therapeutics can be derived. Study design: In this work, we studied the culture of rabbit nucleus pulposus (NP) cells in a collagen microsphere system and compared their cell morphology and expression of a few potential phenotypic markers with that in monolayer culture. Methods: Specifically, rabbit NP cells isolated from both young and old animals were encapsulated and cultured in collagen microspheres with different monomeric concentrations and with different cell encapsulation density for different period of time. Evaluation on the growth kinetics, the viability, the cell morphology, the expression of Types I and II collagen, glycosaminoglycans (GAGs), and Keratin 19, and the ultrastructure of the fiber meshwork were conducted to compare the microsphere 3D culture system and the traditional monolayer cultures. Results: Nucleus pulposus cells in two-dimensional culture lost the phenotypic expression of Type II collagen and keratin 19 and expressed Type I collagen. In contrast, the 3D collagen microsphere culture system consistently outperformed the traditional monolayer culture in maintaining a round morphology and preserving the phenotypes of NP cells with persistent expression of Type II collagen and Keratin 19. These cells also remodeled the template collagen matrix in the microspheres by depositing new matrices, including collagen Type II and GAGs in a cell seeding density and collagen concentration dependent manner. Conclusions: This study demonstrates the appeal of the 3D collagen microsphere system for NP cell culture over traditional monolayer culture because it preserves the phenotypic characteristics of NP cells. This system also enables the NP cells to remodel the template collagen matrix by depositing new matrices, suggesting an innovative way to reconstitute cell-specific and native tissue-like environment in vitro for future studies on stem cell matrix niche and interactions of NP cell with extrinsic factors. © 2011 Elsevier Inc. All rights reserved.en_HK
dc.languageengen_US
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/spineeen_HK
dc.relation.ispartofSpine Journalen_HK
dc.subject3D cultureen_HK
dc.subjectCollagenen_HK
dc.subjectIntervertebral discen_HK
dc.subjectMicroencapsulationen_HK
dc.subjectNucleus pulposus cellsen_HK
dc.subject.meshCell Culture Techniques - methods-
dc.subject.meshCells, Cultured-
dc.subject.meshCollagen - metabolism-
dc.subject.meshIntervertebral Disc - cytology-
dc.subject.meshMicrospheres-
dc.titleThree-dimensional culture of rabbit nucleus pulposus cells in collagen microspheresen_HK
dc.typeArticleen_HK
dc.identifier.emailChan, BP:bpchan@hkucc.hku.hken_HK
dc.identifier.authorityChan, BP=rp00087en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.spinee.2011.07.004en_HK
dc.identifier.pmid21843975-
dc.identifier.scopuseid_2-s2.0-80054091470en_HK
dc.identifier.hkuros196470en_US
dc.identifier.hkuros234176-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-80054091470&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume11en_HK
dc.identifier.issue10en_HK
dc.identifier.spage947en_HK
dc.identifier.epage960en_HK
dc.identifier.isiWOS:000296508700009-
dc.publisher.placeNetherlandsen_HK
dc.relation.projectBioengineering a Prototype Intervertebral Disc Motion Segment-
dc.identifier.scopusauthoridYuan, M=53986920400en_HK
dc.identifier.scopusauthoridLeong, KW=7201577196en_HK
dc.identifier.scopusauthoridChan, BP=7201530390en_HK
dc.identifier.citeulike9694423-

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