Publication: Development and phenotypic characterization of
a high density in vitro model of auricular chondrocytes
with applications in reconstructive plastic surgery
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Date
2006
Authors
Haisch, A. ; Marzahn, U. ; Mobasheri, A. ; Schulze-Tanzil, G. ; Shakibaei, M.
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
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 ß1-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 ß1-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.
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