Please use this identifier to cite or link to this item: http://dx.doi.org/10.14279/depositonce-8009
Main Title: Development of a High-Throughput Screening Assay Based on the 3-Dimensional Pannus Model for Rheumatoid Arthritis
Author(s): Ibold, Yvonne
Frauenschuh, Simone
Kaps, Christian
Sittinger, Michael
Ringe, Jochen
Goetz, Peter M.
Type: Article
Language Code: en
Abstract: The 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)
URI: https://depositonce.tu-berlin.de//handle/11303/8880
http://dx.doi.org/10.14279/depositonce-8009
Issue Date: 2007
Date Available: 8-Jan-2019
DDC Class: 570 Biowissenschaften; Biologie
Subject(s): 3-D culture
interactive co-culture
HTS assay
automation
pannus model
Sponsor/Funder: BMBF, 0313604A, Verbundprojekt: Evaluierung eines interagierenden 3D Testsystems als Krankheitsmodell der rheumatoiden Arthritis (in vitro Pannus Modell) zur effektiven Prüfung von Wirkstoffen, Teilprojekt 1
BMBF, 0313604B, Verbundprojekt: Entwicklung eines interagierenden 3D Testsystems als Krankheitsmodell der rheumatoiden Arthritis (in vitro Pannus Modell) zur effektiven Prüfung von Wirkstoffen, Teilprojekt 2
License: http://rightsstatements.org/vocab/InC/1.0/
Journal Title: Journal of Biomolecular Screening
Publisher: SAGE Publications
Publisher Place: Washington, DC
Volume: 12
Issue: 7
Publisher DOI: 10.1177/1087057107307147
Page Start: 956
Page End: 965
EISSN: 1552-454X
ISSN: 1087-0571
Notes: Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG geförderten) Allianz- bzw. Nationallizenz frei zugänglich.
This 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.
Appears in Collections:FG Bioverfahrenstechnik » Publications

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