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Conference Paper: Intervertebral Disc Cryopreservation

TitleIntervertebral Disc Cryopreservation
Authors
Issue Date2008
PublisherInternational Society of Orthopaedic Surgery and Traumatology.
Citation
SICOT/SIROT 2008 XXIV Triennial World Congress, Hong Kong, 24-28 August 2008, p. abstract no. 17964 How to Cite?
AbstractCryopreserved allogeneic intervertebral disc (IVD) transplantation has been performed clinically for treating disc degeneration. However, the transplanted allograft has undergone progressive degeneration 1. Conventional disc cryopreservation method has been shown to greatly decrease the disc cell viability and hence may be a cause of the degeneration. In this study, we aim to search for a freeze-thaw protocol that is able to retain a maximal cell viability and metabolic activity in the cryopreserved IVD. Porcine lumbar IVDs with endplates were harvested and cryopreserved in various freezing medium 1) 45% Cryo-protective agents (CPAs) (DMSO (dimethylsulfoxide) + propylene glycol), 2) 20% CPAs (DMSO + glycerol), 3) 10% DMSO with synthetic ice-blocker , 4) 10% DMSO, or 5) without CPA. IVDs were incubated at 4°C for 1 hour, freezed at -80°C overnight, and then stored in liquid nitrogen. The frozen IVDs were thawed quickly at 37°C for analysis. Cell prol iferation and proteoglycan production rate were evaluated using Alarma blue and 35S-sulphate incorporation respectively. The distribution of live and dead cells was determined microscopically using a LIVD/DEAD staining. IVD cryopreserved in freezing medium composed of 45% CPAs showed higher disc cell viability compared with that in 10% DMSO. Cell viability was 50% of the fresh IVD tissue when cryopreserved in 45% CPAs but merely 20% when cryopreserved with 10% DMSO. Combination of DMSO and propylene glycol as CPAs for IVD cryopreservation has an advantage over conventional cryopreservation protocol in maintaining disc cell viability. Reference: 1. Ruan D, et al. Lancet 2007; 369:993-9.
DescriptionSession: Spine: basic science
Oral presentation
Persistent Identifierhttp://hdl.handle.net/10722/61622

 

DC FieldValueLanguage
dc.contributor.authorChan, CWen_HK
dc.contributor.authorLeung, YLen_HK
dc.contributor.authorLu, WWen_HK
dc.contributor.authorChan, Den_HK
dc.contributor.authorLuk, KDKen_HK
dc.contributor.authorCheung, KMCen_HK
dc.date.accessioned2010-07-13T03:43:40Z-
dc.date.available2010-07-13T03:43:40Z-
dc.date.issued2008en_HK
dc.identifier.citationSICOT/SIROT 2008 XXIV Triennial World Congress, Hong Kong, 24-28 August 2008, p. abstract no. 17964-
dc.identifier.urihttp://hdl.handle.net/10722/61622-
dc.descriptionSession: Spine: basic science-
dc.descriptionOral presentation-
dc.description.abstractCryopreserved allogeneic intervertebral disc (IVD) transplantation has been performed clinically for treating disc degeneration. However, the transplanted allograft has undergone progressive degeneration 1. Conventional disc cryopreservation method has been shown to greatly decrease the disc cell viability and hence may be a cause of the degeneration. In this study, we aim to search for a freeze-thaw protocol that is able to retain a maximal cell viability and metabolic activity in the cryopreserved IVD. Porcine lumbar IVDs with endplates were harvested and cryopreserved in various freezing medium 1) 45% Cryo-protective agents (CPAs) (DMSO (dimethylsulfoxide) + propylene glycol), 2) 20% CPAs (DMSO + glycerol), 3) 10% DMSO with synthetic ice-blocker , 4) 10% DMSO, or 5) without CPA. IVDs were incubated at 4°C for 1 hour, freezed at -80°C overnight, and then stored in liquid nitrogen. The frozen IVDs were thawed quickly at 37°C for analysis. Cell prol iferation and proteoglycan production rate were evaluated using Alarma blue and 35S-sulphate incorporation respectively. The distribution of live and dead cells was determined microscopically using a LIVD/DEAD staining. IVD cryopreserved in freezing medium composed of 45% CPAs showed higher disc cell viability compared with that in 10% DMSO. Cell viability was 50% of the fresh IVD tissue when cryopreserved in 45% CPAs but merely 20% when cryopreserved with 10% DMSO. Combination of DMSO and propylene glycol as CPAs for IVD cryopreservation has an advantage over conventional cryopreservation protocol in maintaining disc cell viability. Reference: 1. Ruan D, et al. Lancet 2007; 369:993-9.-
dc.languageengen_HK
dc.publisherInternational Society of Orthopaedic Surgery and Traumatology.-
dc.relation.ispartofSICOT/SIROT World Congress-
dc.titleIntervertebral Disc Cryopreservationen_HK
dc.typeConference_Paperen_HK
dc.identifier.emailChan, CW: samanthacn@gmail.comen_HK
dc.identifier.emailLeung, YL: vicleung@hku.hken_HK
dc.identifier.emailLu, WW: wwlu@hkusua.hku.hken_HK
dc.identifier.emailChan, D: chand@hku.hken_HK
dc.identifier.emailLuk, KDK: hrmoldk@hkucc.hku.hken_HK
dc.identifier.emailCheung, KMC: cheungmc@hku.hken_HK
dc.identifier.authorityLeung, YL=rp01764en_HK
dc.identifier.authorityLu, WW=rp00411en_HK
dc.identifier.authorityChan, D=rp00540en_HK
dc.identifier.hkuros166115en_HK
dc.identifier.spageabstract no. 17964-
dc.identifier.epageabstract no. 17964-
dc.publisher.placeFrance-

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