Optimal 3D Culture of Primary Articular Chondrocytes for Use in the Rotating Wall Vessel Bioreactor
| Autor: | Lindsey W. Catlin, Liliana F. Mellor, Travis L. Baker, Julia Thom Oxford, Raquel J. Brown |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Primary (chemistry)
Rotation Chemistry Cell Culture Techniques Public Health Environmental and Occupational Health Monolayer culture Gene Expression Microcarrier Osteoarthritis medicine.disease Article Cartilage tissue engineering Bioreactors Chondrocytes Microscopy Fluorescence Cell culture medicine Bioreactor Animals Cattle Cellular Morphology Biomedical engineering |
| Zdroj: | Aviation, Space, and Environmental Medicine. 85:798-804 |
| ISSN: | 0095-6562 |
| DOI: | 10.3357/asem.3905.2014 |
| Popis: | Reliable culturing methods for primary articular chon-drocytes are essential to study the effects of loading and unloading on joint tissue at the cellular level. Due to the limited proliferation capacity of primary chondrocytes and their tendency to dedifferentiate in conven-tional culture conditions, long-term culturing conditions of primary chondrocytes can be challenging. The goal of this study was to develop a suspension culturing technique that not only would retain the cellular morphology, but also maintain gene expression characteristics of pri-mary articular chondrocytes. Methods: Three-dimensional culturing methods were compared and optimized for primary articular chondro-cytes in the rotating wall vessel bioreactor, which changes the mechani-cal culture conditions to provide a form of suspension culture optimized for low shear and turbulence. We performed gene expression analysis and morphological characterization of cells cultured in alginate beads, Cytopore-2 microcarriers, primary monolayer culture, and passaged monolayer cultures using reverse transcription-PCR and laser scanning confocal microscopy. Results: Primary chondrocytes grown on Cytopore-2 microcarriers maintained the phenotypical morphology and gene ex-pression pattern observed in primary bovine articular chondrocytes, and retained these characteristics for up to 9 d. Discussion: Our results pro-vide a novel and alternative culturing technique for primary chondro-cytes suitable for studies that require suspension such as those using the rotating wall vessel bioreactor. In addition, we provide an alternative culturing technique for primary chondrocytes that can impact future mechanistic studies of osteoarthritis progression, treatments for cartil-age damage and repair, and cartilage tissue engineering. Keywords |
| Databáze: | OpenAIRE |
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