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Development and phenotypic characterization of a high density in vitro model of auricular chondrocytes with applications in reconstructive plastic surgery.

Haisch, A, Marzahn, U, Mobasheri, A, Schulze-Tanzil, G and Shakibaei, M (2006) Development and phenotypic characterization of a high density in vitro model of auricular chondrocytes with applications in reconstructive plastic surgery. Histol Histopathol, 21 (5). pp. 467-476.

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Cultivation of phenotypically stable auricular chondrocytes will have applications in autologous chondrocyte transplantation and reconstructive surgery of cartilage. Chondrocytes grown in monolayer culture rapidly dedifferentiate assuming a fibroblast-like morphology and lose their cartilage-specific pattern of gene expression. Three-dimensional high-density culture models mimic more closely the in vivo conditions of cartilage. Therefore, this study was undertaken to test whether the high-density cultures might serve as a suitable model system to acquire phenotypically and functionally differentiated auricular chondrocytes from porcine cartilage. Freshly isolated porcine auricular chondrocytes were cultured for 7 passages in monolayer culture. From each passage (passage 0 and 1-7) cells were introduced to high-density cultures and examined by transmission electron microscopy. Western blotting was used to analyse the expression of cartilage-specific markers, such as collagen type II and cartilage specific proteoglycan, fibronectin, cell adhesion and signal transduction receptor beta1-integrin, matrix metalloproteinases (MMP-9, MMP-13), cyclo-oxygenase (COX)-2 and the apoptosis commitment marker, activated caspase-3. When dedifferentiated auricular chondrocytes from monolayer passages 0-4 were cultured in high-density culture, they recovered their chondrocytic phenotype and formed cartilage nodules surrounded by fibroblast-like cells and synthesised collagen type II, proteoglycans, fibronectin and beta1-integrins. However, chondrocytes from monolayer passages 5-7 did not redifferentiate to chondrocytes even when transferred to high-density culture, and did not synthesize a chondrocyte-specific extracellular matrix. Instead, they produced increasing amounts of MMP-9, MMP-13, COX-2, activated caspase-3 and underwent apoptosis. Three-dimensional high-density cultures may therefore be used to obtain sufficient quantities of fully differentiated auricular chondrocytes for autologous chondrocyte transplantation and reconstructive plastic surgery.

Item Type: Article
Divisions : Surrey research (other units)
Authors :
Haisch, A
Marzahn, U
Schulze-Tanzil, G
Shakibaei, M
Date : May 2006
Uncontrolled Keywords : Animals, Antigens, CD29, Blotting, Western, Caspase 3, Caspases, Cell Count, Cell Culture Techniques, Cell Differentiation, Cell Proliferation, Cell Transplantation, Cells, Cultured, Chondrocytes, Collagen Type II, Collagenases, Ear, Fibronectins, Matrix Metalloproteinase 13, Matrix Metalloproteinase 9, Microscopy, Electron, Transmission, Phenotype, Proteoglycans, Reconstructive Surgical Procedures, Surgery, Plastic, Swine
Related URLs :
Depositing User : Symplectic Elements
Date Deposited : 17 May 2017 10:12
Last Modified : 24 Jan 2020 18:43

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