title: A Luciferase-Based Quick Potency Assay to Predict Chondrogenic Differentiation.
abstract: Chondrogenic differentiation of adipose-derived stem cells (ASC) is challenging but highly promising for cartilage repair. Large donor variability of chondrogenic differentiation potential raises the risk for transplantation of cells with reduced efficacy and a low chondrogenic potential. Therefore, quick potency assays are required to control the potency of the isolated cells before cell transplantation. Current in vitro methods to analyze the differentiation capacity are time-consuming, and thus, a novel enhancer and tissue-specific promoter combination was used for the detection of chondrogenic differentiation of ASC in a novel quick potency bioassay. Human primary ASC were cotransfected with the Metridia luciferase-based collagen type II reporter gene pCMVE_ACDCII-MetLuc together with a Renilla control plasmid and analyzed for their chondrogenic potential. On day 3 after chondrogenic induction, the luciferase activity was induced in all tested donors under three-dimensional culture conditions and, in a second approach, also under two-dimensional (2D) culture conditions. With our newly developed quick potency bioassay, we can determine chondrogenic potential already after 3 days of chondrogenic induction and under 2D culture conditions. This will enhance the efficiency of testing cell functionality, which should allow in the future to predict the suitability of cells derived from individual patients for cell therapies in a very short time and at low costs.
authors.affiliation: 1 Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center , Linz/Vienna, Austria .|2 Austrian Cluster for Tissue Regeneration , Vienna, Austria .
authors.forename: Eleni
authors.initials: E
authors.lastname: Oberbauer
authors.affiliation: 1 Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center , Linz/Vienna, Austria .|2 Austrian Cluster for Tissue Regeneration , Vienna, Austria .
authors.forename: Carolin
authors.initials: C
authors.lastname: Steffenhagen
authors.affiliation: 1 Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center , Linz/Vienna, Austria .|2 Austrian Cluster for Tissue Regeneration , Vienna, Austria .|3 Biomaterials and Tissue Engineering Group, Department of Oral Biology, School of Dentistry, University of Leeds , Leeds, United Kingdom .
authors.forename: Georg
authors.initials: G
authors.lastname: Feichtinger
authors.affiliation: 1 Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center , Linz/Vienna, Austria .|2 Austrian Cluster for Tissue Regeneration , Vienna, Austria .
authors.forename: Florian
authors.initials: F
authors.lastname: Hildner
authors.affiliation: 1 Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center , Linz/Vienna, Austria .|2 Austrian Cluster for Tissue Regeneration , Vienna, Austria .
authors.forename: Ara
authors.initials: A
authors.lastname: Hacobian
authors.affiliation: 1 Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center , Linz/Vienna, Austria .|2 Austrian Cluster for Tissue Regeneration , Vienna, Austria .|4 Red Cross Blood Transfusion Service of Upper Austria, Linz, Austria .
authors.forename: Martin
authors.initials: M
authors.lastname: Danzer
authors.affiliation: 1 Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center , Linz/Vienna, Austria .|2 Austrian Cluster for Tissue Regeneration , Vienna, Austria .|4 Red Cross Blood Transfusion Service of Upper Austria, Linz, Austria .
authors.forename: Christian
authors.initials: C
authors.lastname: Gabriel
authors.affiliation: 1 Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center , Linz/Vienna, Austria .|2 Austrian Cluster for Tissue Regeneration , Vienna, Austria .
authors.forename: Heinz
authors.initials: H
authors.lastname: Redl
authors.affiliation: 1 Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center , Linz/Vienna, Austria .|2 Austrian Cluster for Tissue Regeneration , Vienna, Austria .
authors.forename: Susanne
authors.initials: S
authors.lastname: Wolbank
mesh_terms: D000273:Adipose Tissue / Q000166:cytology* / Q000378:metabolism; D001681:Biological Assay / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism; D020219:Chondrogenesis / Q000502:physiology*; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods
chemical_list: D024043:Collagen Type II; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D008156:Luciferases