42 research outputs found
Development and phenotypic characterization of a high density in vitro model of auricular chondrocytes with applications in reconstructive plastic surgery
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 β1-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 β1-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
Development and phenotypic characterization of a high density in vitro model of auricular chondrocytes with applications in reconstructive plastic surgery
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
Development and phenotypic characterization of a high density in vitro model of auricular chondrocytes with applications in reconstructive plastic surgery.
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