Adenoviral transduction supports matrix expression of alginate cultured articular chondrocytes
| Autor: | Diana Pohle, R. Kasch, Thomas Mittlmeier, Rainer Bader, Brigitte Müller-Hilke, Philipp Herlyn, B.M. Pützer |
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| Rok vydání: | 2012 |
| Předmět: |
Cartilage
Articular Genetic Markers Male Alginates Cell Culture Techniques Bioengineering Transfection Polymerase Chain Reaction Applied Microbiology and Biotechnology Statistics Nonparametric Chondrocyte Adenoviridae Extracellular matrix Transduction (genetics) Chondrocytes Glucuronic Acid Transduction Genetic Gene expression medicine Humans Cells Cultured Aged Aged 80 and over Regulation of gene expression Extracellular Matrix Proteins biology Chemistry Hexuronic Acids Cartilage Femur Head Middle Aged Cell biology Transplantation medicine.anatomical_structure Gene Expression Regulation Proteoglycan Immunology biology.protein Female Biotechnology |
| Zdroj: | Biotechnology and Bioengineering. 109:2402-2408 |
| ISSN: | 0006-3592 |
| Popis: | The present study examines the effects of adenoviral (Ad) transduction of human primary chondrocyte on transgene expression and matrix production. Primary chondrocytes were isolated from healthy articular cartilage and from cartilage with mild osteoarthritis (OA), transduced with an Ad vector and either immediately cultured in alginate or expanded in monolayer before alginate culture. Proteoglycan production was measured using dimethylmethylene blue (DMMB) assay and matrix gene expression was quantified by real-time PCR. Viral infection of primary chondrocytes results in a stable long time transgene expression for up to 13 weeks. Ad transduction does not significantly alter gene expression and matrix production if chondrocytes are immediately embedded in alginate. However, if expanded prior to three dimension (3D) culture in alginate, chondrocytes produce not only more proteoglycans compared to non-transduced controls, but also display an increased anabolic and decreased catabolic activity compared to non-transduced controls. We therefore suggest that successful autologous chondrocyte transplantation (ACT) should combine adenoviral transduction of primary chondrocytes with expansion in monolayer followed by 3D culture. Future studies will be needed to investigate whether the subsequent matrix production can be further improved by using Ad vectors bearing genes encoding matrix proteins. |
| Databáze: | OpenAIRE |
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