Phenotypic analysis of bovine chondrocytes cultured in 3D collagen sponges: effect of serum substitutes

Yates, Karen E.; Allemann, Florin; Glowacki, Julie

doi:10.1007/s10561-005-5810-0

Cited by 51 publications

(33 citation statements)

References 42 publications

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“…23 It is well known that in monolayer 2D culture, chondrocytes dedifferentiate to a less specialized fibroblastic phenotype and produce less cartilage ECM. When grown in 3D porous collagen sponges, however, production of cartilage-specific ECM 20 and expression of chondrocyte genes (aggrecan core protein and collagen type II) 24 were maintained with time, in contrast to the decreases seen in monolayer.…”

Section: In Vitro Histogenesismentioning

confidence: 87%

Collagen scaffolds for tissue engineering

2007

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There are two major approaches to tissue engineering for regeneration of tissues and organs. One involves cell‐free materials and/or factors and one involves delivering cells to contribute to the regeneraion process. Of the many scaffold materials being investigated, collagen type I, with selective removal of its telopeptides, has been shown to have many advantageous features for both of these approaches. Highly porous collagen lattice sponges have been used to support in vitro growth of many types of tissues. Use of bioreactors to control in vitro perfusion of medium and to apply hydrostatic fluid pressure has been shown to enhance histogenesis in collagen scaffolds. Collagen sponges have also been developed to contain differentiating‐inducing materials like demineralized bone to stimulate differentiation of cartilage tissue both in vitro and in vivo. © 2007 Wiley Periodicals, Inc. Biopolymers 89: 338–344, 2008.This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

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Section: In Vitro Histogenesismentioning

confidence: 87%

Collagen scaffolds for tissue engineering

2007

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“…Complementary deoxyribonucleic acid (DNA) was synthesized using SuperScript II (Invitrogen Life Technologies, Carlsbad, Calif., USA), following the manufacturer's instruction. Real-time RT-PCR was performed using SYBR Green Realtime PCR Master Mix Plus (Toyobo, Osaka, Japan) with primers of aggrecan core protein (Agg-core) and collagen type II ( Col-2 ) ( table 1 ) [Yates et al, 2005;Mio et al, 2007]. Amplifications of the complementary DNA samples were assessed in triplicate in 96-well plates at a final volume of 20 μl at 50 PCR cycles, consisting of a denaturation step at 95 ° C for 15 s and an annealing/extension step at 60 ° C for 1 min.…”

Section: Molecular Evaluationmentioning

confidence: 99%

Off-Loading of Cyclic Hydrostatic Pressure Promotes Production of Extracellular Matrix by Chondrocytes

Tatsumura

Sakane

Ochiai

et al. 2013

Cells Tissues Organs

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The addition of cyclic hydrostatic pressure (cHP) to cell culture medium has been used to promote extracellular matrix (ECM) production by articular chondrocytes. Though a combination of cHP followed by atmospheric pressure (AP) has been examined previously, the rationale of such a combination was unclear. We compared the effects of loading once versus twice (combinations of cHP followed by AP) regarding both gene expression and biochemical and histological phenotypes of chondrocytes. Isolated bovine articular chondrocytes were embedded in a collagen gel and incubated for 14 days under conditions combining cHP and AP. The gene expression of aggrecan core protein and collagen type II were upregulated in response to cHP, and those levels were maintained for at least 4 days after cHP treatment. Accumulation of cartilage-specific sulfated glycosaminoglycans following cHP for 7 days and subsequent AP for 7 days was significantly greater than that of the AP control (p < 0.05). Therefore, incubation at AP after loading with cHP was found to beneficially affect ECM accumulation. Manipulating algorithms of cHP combined with AP will be useful in producing autologous chondrocyte-based cell constructs for implantation.

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“…In all tested scaffolds, transcript levels of cartilage-related genes, like Col 2, Agr, Sox 9, were upregulated. [27][28][29][30][31][32] Two different types of hydrogels were examined in the present study: alginate and alginate/agarose. A 14-day incubation period was selected to allow for sufficient redifferentiation.…”

Section: Human Chondrocyte Gene Expression In Vitromentioning

confidence: 99%

Quantitative analysis of gene expression in human articular chondrocytes assigned for autologous implantation

Barlič

Drobnič

Maličev

et al. 2008

Journal Orthopaedic Research

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Autologous chondrocyte implantation (ACI) relies on the implantation of in vitro expanded cells. The aim was to study the dedifferentiation of human articular chondrocytes under different cultivating conditions [days 0-10 in the primary culture (P0); passages in a monolayer from P0 to P3; monolayer vs. alginate and monolayer vs. alginate/agarose hydrogels] using real-time PCR analysis. The relative gene expressions for collagen type I and II, aggrecan and versican were quantified and the corresponding differentiation indexes (Col2/Col1, Agr/Ver) were calculated. The values of both differentiation indexes decreased exponentially with time in the P0 monolayer culture, and continued with a significant decrease over the subsequent monolayer passages. On the contrary, the chondrocytes seeded in either of the hydrogels significantly increased the indexes compared to their parallel monolayer cultures. These results indicate that alginate and alginate/agarose hydrogels offer an appropriate environment for human articular chondrocytes to redifferentiate after being expanded in vitro. Therefore the three-dimensional (3D) hydrogel chondrocyte cultures present not only surgical, but also biological advantage over the classic suspension-periosteum chondrocyte implantation. ß

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Phenotypic analysis of bovine chondrocytes cultured in 3D collagen sponges: effect of serum substitutes

Cited by 51 publications

References 42 publications

Collagen scaffolds for tissue engineering

Collagen scaffolds for tissue engineering

Off-Loading of Cyclic Hydrostatic Pressure Promotes Production of Extracellular Matrix by Chondrocytes

Quantitative analysis of gene expression in human articular chondrocytes assigned for autologous implantation

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