Development of a High-Throughput Screening Assay Based on the 3-Dimensional Pannus Model for Rheumatoid Arthritis

dc.contributor.authorIbold, Yvonne
dc.contributor.authorFrauenschuh, Simone
dc.contributor.authorKaps, Christian
dc.contributor.authorSittinger, Michael
dc.contributor.authorRinge, Jochen
dc.contributor.authorGoetz, Peter M.
dc.date.accessioned2019-01-08T17:37:59Z
dc.date.available2019-01-08T17:37:59Z
dc.date.issued2007
dc.descriptionDieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG geförderten) Allianz- bzw. Nationallizenz frei zugänglich.de
dc.descriptionThis publication is with permission of the rights owner freely accessible due to an Alliance licence and a national licence (funded by the DFG, German Research Foundation) respectively.en
dc.description.abstractThe 3-dimensional (3-D) pannus model for rheumatoid arthritis (RA) is based on the interactive co-culture of cartilage and synovial fibroblasts (SFs). Besides the investigation of the pathogenesis of RA, it can be used to analyze the active profiles of antirheumatic pharmaceuticals and other bioactive substances under in vitro conditions. For a potential application in the industrial drug-screening process as a transitional step between 2-dimensional (2-D) cell-based assays and in vivo animal studies, the pannus model was developed into an in vitro high-throughput screening (HTS) assay. Using the CyBi™-Disk workstation for parallel liquid handling, the main cell culture steps of cell seeding and cultivation were automated. Chondrocytes were isolated from articular cartilage and seeded directly into 96-well microplates in high-density pellets to ensure formation of cartilage-specific extracellular matrix (ECM). Cell seeding was performed automatically and manually to compare both processes regarding accuracy, reproducibility, consistency, and handling time. For automated cultivation of the chondrocyte pellet cultures, a sequential program was developed using the CyBio Control software to minimize shear forces and handling time. After 14 days of cultivation, the pannus model was completed by coating the cartilage pellets with a layer of human SFs. The effects due to automation in comparison to manual handling were analyzed by optical analysis of the pellets, histological and immunohistochemical staining, and real-time PCR. Automation of this in vitro model was successfully achieved and resulted in an improved quality of the generated pannus cultures by enhancing the formation of cartilage-specific ECM. In addition, automated cell seeding and media exchange increased the efficiency due to a reduction of labor intensity and handling time. (Journal of Biomolecular Screening 2007:956-965)en
dc.description.sponsorshipBMBF, 0313604A, Verbundprojekt: Evaluierung eines interagierenden 3D Testsystems als Krankheitsmodell der rheumatoiden Arthritis (in vitro Pannus Modell) zur effektiven Prüfung von Wirkstoffen, Teilprojekt 1en
dc.description.sponsorshipBMBF, 0313604B, Verbundprojekt: Entwicklung eines interagierenden 3D Testsystems als Krankheitsmodell der rheumatoiden Arthritis (in vitro Pannus Modell) zur effektiven Prüfung von Wirkstoffen, Teilprojekt 2en
dc.identifier.eissn1552-454X
dc.identifier.issn1087-0571
dc.identifier.urihttps://depositonce.tu-berlin.de/handle/11303/8880
dc.identifier.urihttp://dx.doi.org/10.14279/depositonce-8009
dc.language.isoen
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/
dc.subject.ddc570 Biowissenschaften; Biologiede
dc.subject.other3-D cultureen
dc.subject.otherinteractive co-cultureen
dc.subject.otherHTS assayen
dc.subject.otherautomationen
dc.subject.otherpannus modelen
dc.titleDevelopment of a High-Throughput Screening Assay Based on the 3-Dimensional Pannus Model for Rheumatoid Arthritisen
dc.typeArticleen
dc.type.versionpublishedVersionen
dcterms.bibliographicCitation.doi10.1177/1087057107307147
dcterms.bibliographicCitation.issue7
dcterms.bibliographicCitation.journaltitleJournal of Biomolecular Screeningen
dcterms.bibliographicCitation.originalpublishernameSAGE Publicationsen
dcterms.bibliographicCitation.originalpublisherplaceWashington, DCen
dcterms.bibliographicCitation.pageend965
dcterms.bibliographicCitation.pagestart956
dcterms.bibliographicCitation.volume12
tub.accessrights.dnbdomain
tub.affiliationFak. 3 Prozesswissenschaften::Inst. Biotechnologie::FG Bioverfahrenstechnikde
tub.affiliation.facultyFak. 3 Prozesswissenschaftende
tub.affiliation.groupFG Bioverfahrenstechnikde
tub.affiliation.instituteInst. Biotechnologiede
tub.publisher.universityorinstitutionTechnische Universität Berlinde

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