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Article: Cryopreserved intervertebral disc with injected bone marrow-derived stromal cells: a feasibility study using organ culture

TitleCryopreserved intervertebral disc with injected bone marrow-derived stromal cells: a feasibility study using organ culture
Authors
KeywordsBone marrow-derived mesenchymal stromal cells
Bovine
Cell viability
Cryopreservation
Gene expression
Intervertebral disc
Issue Date2010
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/spinee
Citation
Spine Journal, 2010, v. 10 n. 6, p. 486-496 How to Cite?
AbstractBackground Context: A recent clinical study demonstrated that cryopreserved allogeneic intervertebral disc transplantation relieved pain and preserved motion, thus opening up a new treatment option for degenerative disc disease. However, these transplanted discs continued to degenerate, possibly due to a lack of viable cells. Bone marrow-derived stromal cell (BMSC) implantation has been shown to delay disc degeneration. Purpose: This study examined the viability over time of endogenous and injected BMSCs in cryopreserved disc under simulated-physiological loading conditions. Study Design/ Setting: An in vitro study of BMSCs injected into cryopreserved bovine caudal discs. Methods: Bovine caudal discs were harvested and cryopreserved at -196°C. After thawing, PKH-26-labeled BMSCs embedded in peptide hydrogel carrier were injected into the nucleus pulposus. Two BMSC injection quantities, that is, 1×10 5 and 2.5×10 5 were examined. Discs with injected cells were maintained in a bioreactor for 7 days under simulated-physiological loading. Cell viability (staining), gene expression (reverse transcription-polymerase chain reaction) profile, and proteoglycan content (histologically) were evaluated. Results: Forty percent of endogenous cell viability was maintained after freeze thawing. Over the 7-day culture, this did not change further. However, there was upregulation of Col1a2 and Mmp-13 and downregulation of Col2a1gene expression. Sixty percent of BMSCs survived the initial injection procedure, and only 20% remained alive after 7 days of culture. Bone marrow-derived stromal cell implantation did not alter the viability of the endogenous cells, but discs injected with 1×105 BMSCs showed significantly higher ACAN expression than sham discs. Conclusions: Although only 40% of cells survived cryopreservation, these endogeneous cells continued to survive over 7 days if maintained under simulated-physiological loading conditions. Although only a small portion of injected BMSCs survived, they did have some effect on the matrix protein gene expression profile. Their influence on native cells requires long-term evaluation. © 2010 Elsevier Inc. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/129091
ISSN
2015 Impact Factor: 2.66
2015 SCImago Journal Rankings: 1.153
ISI Accession Number ID
Funding AgencyGrant Number
KMCC
AO Foundation
Canton of Grison
Funding Information:

KMCC (research support for staff and materials, Synthes).

References

 

DC FieldValueLanguage
dc.contributor.authorChan, SCWen_HK
dc.contributor.authorGantenbeinRitter, Ben_HK
dc.contributor.authorLeung, VYLen_HK
dc.contributor.authorChan, Den_HK
dc.contributor.authorCheung, KMCen_HK
dc.contributor.authorIto, Ken_HK
dc.date.accessioned2010-12-23T08:32:23Z-
dc.date.available2010-12-23T08:32:23Z-
dc.date.issued2010en_HK
dc.identifier.citationSpine Journal, 2010, v. 10 n. 6, p. 486-496en_HK
dc.identifier.issn1529-9430en_HK
dc.identifier.urihttp://hdl.handle.net/10722/129091-
dc.description.abstractBackground Context: A recent clinical study demonstrated that cryopreserved allogeneic intervertebral disc transplantation relieved pain and preserved motion, thus opening up a new treatment option for degenerative disc disease. However, these transplanted discs continued to degenerate, possibly due to a lack of viable cells. Bone marrow-derived stromal cell (BMSC) implantation has been shown to delay disc degeneration. Purpose: This study examined the viability over time of endogenous and injected BMSCs in cryopreserved disc under simulated-physiological loading conditions. Study Design/ Setting: An in vitro study of BMSCs injected into cryopreserved bovine caudal discs. Methods: Bovine caudal discs were harvested and cryopreserved at -196°C. After thawing, PKH-26-labeled BMSCs embedded in peptide hydrogel carrier were injected into the nucleus pulposus. Two BMSC injection quantities, that is, 1×10 5 and 2.5×10 5 were examined. Discs with injected cells were maintained in a bioreactor for 7 days under simulated-physiological loading. Cell viability (staining), gene expression (reverse transcription-polymerase chain reaction) profile, and proteoglycan content (histologically) were evaluated. Results: Forty percent of endogenous cell viability was maintained after freeze thawing. Over the 7-day culture, this did not change further. However, there was upregulation of Col1a2 and Mmp-13 and downregulation of Col2a1gene expression. Sixty percent of BMSCs survived the initial injection procedure, and only 20% remained alive after 7 days of culture. Bone marrow-derived stromal cell implantation did not alter the viability of the endogenous cells, but discs injected with 1×105 BMSCs showed significantly higher ACAN expression than sham discs. Conclusions: Although only 40% of cells survived cryopreservation, these endogeneous cells continued to survive over 7 days if maintained under simulated-physiological loading conditions. Although only a small portion of injected BMSCs survived, they did have some effect on the matrix protein gene expression profile. Their influence on native cells requires long-term evaluation. © 2010 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.subjectBone marrow-derived mesenchymal stromal cellsen_HK
dc.subjectBovineen_HK
dc.subjectCell viabilityen_HK
dc.subjectCryopreservationen_HK
dc.subjectGene expressionen_HK
dc.subjectIntervertebral discen_HK
dc.subject.meshBone Marrow Cells - cytology - metabolism-
dc.subject.meshBone Marrow Transplantation-
dc.subject.meshCryopreservation-
dc.subject.meshIntervertebral Disk-
dc.subject.meshStromal Cells - cytology - metabolism-
dc.titleCryopreserved intervertebral disc with injected bone marrow-derived stromal cells: a feasibility study using organ cultureen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1529-9430&volume=10&issue=6&spage=486–496&epage=&date=2010&atitle=Cryopreserved+intervertebral+disc+with+injected+bone+marrow-derived+stromal+cells:+a+feasibility+study+using+organ+culture-
dc.identifier.emailLeung, VYL: vicleung@hku.hken_HK
dc.identifier.emailChan, D: chand@hkucc.hku.hken_HK
dc.identifier.emailCheung, KMC: cheungmc@hku.hken_HK
dc.identifier.authorityLeung, VYL=rp01764en_HK
dc.identifier.authorityChan, D=rp00540en_HK
dc.identifier.authorityCheung, KMC=rp00387en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.spinee.2009.12.019en_HK
dc.identifier.pmid20171933-
dc.identifier.scopuseid_2-s2.0-77952420722en_HK
dc.identifier.hkuros178100en_US
dc.identifier.hkuros180119-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-77952420722&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume10en_HK
dc.identifier.issue6en_HK
dc.identifier.spage486en_HK
dc.identifier.epage496en_HK
dc.identifier.isiWOS:000278797400004-
dc.publisher.placeNetherlandsen_HK
dc.identifier.scopusauthoridChan, SCW=54970852000en_HK
dc.identifier.scopusauthoridGantenbeinRitter, B=25824884300en_HK
dc.identifier.scopusauthoridLeung, VYL=35337438900en_HK
dc.identifier.scopusauthoridChan, D=7402216545en_HK
dc.identifier.scopusauthoridCheung, KMC=7402406754en_HK
dc.identifier.scopusauthoridIto, K=24537442500en_HK
dc.identifier.citeulike6863779-

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