Three-dimensional culture of human disc cells within agarose or a collagen sponge: assessment of proteoglycan production

Gruber, Helen E.; Hoelscher, Gretchen L.; Leslie, Kelly; Ingram, Jane A.; Hanley, Edward N.

doi:10.1016/j.biomaterials.2005.06.032

Cited by 123 publications

(89 citation statements)

References 17 publications

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“…Finally, cells were encapsulated in 2% agarose disks (8mm in diameter, 1 mm thick) at a density of 2.4×10 7 cells/ml. Three-dimensional agarose culture has been shown to maintain the phenotype of IVD cells [23][24][25]. After an initial 24-hour culture in the highglucose DMEM containing 10% FBS and 1% antibiotic-antimycotic at 37°C, the agarose disks were separated into four groups which were incubated in the DMEM with glucose concentrations of 1, 2.5, 5, and 25 mM for another 24 hours before the oxygen consumption rate measurement.…”

Section: Methodsmentioning

confidence: 99%

Effects of Low Glucose Concentrations on Oxygen Consumption Rates of Intervertebral Disc Cells

Huang¹,

Yuan²,

Jackson³

et al. 2007

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Study Design-Investigation of the effects of low glucose concentrations on the oxygen consumption rates of intervertebral disc cells.Objectives-To determine the oxygen consumption rate of porcine annulus fibrosus (AF) cells at different glucose concentrations and to examine the differences in the oxygen consumption rate between AF and nucleus pulposus (NP) cells at different glucose levels.Summary of Background Data-Poor nutrient supply has been suggested as a potential mechanism for degeneration of the intervertebral disc (IVD). Distribution of nutrients in the IVD is strongly dependent on transport properties of the tissue and cellular metabolic rates. Previous studies have shown dependence of oxygen consumption rate of IVD cells on oxygen tension, pH levels, and glucose levels outside the physiological range. However, the oxygen consumption rate of AF cells at in vivo glucose levels has not been investigated.Methods-IVD cells were isolated from the outer AF and NP of 4-5 month-old porcine lumbar discs. The changes in oxygen tension were recorded when cells were cultured in sealed metabolism chamber. The oxygen consumption rate of cells was determined by theoretical curve fitting using the Michaelis-Menten equation.Results-The outer AF cells cultured in high glucose medium (25mM) exhibited the lowest oxygen consumption rate whereas no significant differences in oxygen consumption rates were found among outer AF cells cultured at physiological glucose levels (i.e., 1 mM, 2.5 mM, 5 mM). The oxygen consumption rate of NP cells was significantly greater than that of outer AF cells.Conclusions-Since the oxygen consumption rates determined in this study are comparable to the findings in the literature, this study has developed a new alternative method for determining oxygen consumption rate. The oxygen consumption rates of IVD cells reported in this study will be valuable for theoretically predicting local oxygen concentrations in IVD, which can provide a better understanding of transport of oxygen in the discs.

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Section: Methodsmentioning

confidence: 99%

Effects of Low Glucose Concentrations on Oxygen Consumption Rates of Intervertebral Disc Cells

Huang¹,

Yuan²,

Jackson³

et al. 2007

Spine

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“…• C. Agarose is commonly used in tissue engineering; Gruber et al (2006) and Jain et al (2006) have noted that it exhibits mechanical properties similar to those of soft tissue and exhibits good cell compatibility. In addition, agarose can be readily patterned into a desired shape.…”

Section: Resultsmentioning

confidence: 99%

Vascular tissue engineering by computer-aided laser micromachining

Doraiswamy

Narayan

2010

Phil. Trans. R. Soc. A.

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Many conventional technologies for fabricating tissue engineering scaffolds are not suitable for fabricating scaffolds with patient-specific attributes. For example, many conventional technologies for fabricating tissue engineering scaffolds do not provide control over overall scaffold geometry or over cell position within the scaffold. In this study, the use of computer-aided laser micromachining to create scaffolds for vascular tissue networks was investigated. Computer-aided laser micromachining was used to construct patterned surfaces in agarose or in silicon, which were used for differential adherence and growth of cells into vascular tissue networks. Concentric three-ring structures were fabricated on agarose hydrogel substrates, in which the inner ring contained human aortic endothelial cells, the middle ring contained HA587 human elastin and the outer ring contained human aortic vascular smooth muscle cells. Basement membrane matrix containing vascular endothelial growth factor and heparin was to promote proliferation of human aortic endothelial cells within the vascular tissue networks. Computer-aided laser micromachining provides a unique approach to fabricate small-diameter blood vessels for bypass surgery as well as other artificial tissues with complex geometries.

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“…Collagen scaffolds also represent promising constructs to maintain functional human disc cells as demonstrated by their high proliferation rate and their rate of proteoglycan synthesis in this type of matrix (Gruber et al, 2006). Nevertheless, it has been reported the proteoglycan content of similar scaffolds cultured up to 60 days never exceeded the 10% of that present in the mature nucleus pulposus.…”

Section: Culturing Chondrogenic Cells In Naturally-derived Hydrogelsmentioning

confidence: 99%

Three-dimensional cultures of osteogenic and chondrogenic cells: A tissue engineering approach to mimic bone and cartilage in vitro

Tortelli

Cancedda²

2009

eCM

103

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Capturing the complexity of bone and cartilage into three-dimensional in vitro models remains one of the most important challenges in the field of the tissue engineering. Indeed, the development and the optimization of novel culture systems may be necessary to face the next questions of bone and cartilage physiology. The models should faithfully mimic these tissues, resembling their organization, their mechanical properties and their physiological response to different stimuli. Here we review the recent advances in the field of the three-dimensional cultures of both osteogenic and chondrogenic cells. In particular, we highlight the most important studies that, to our knowledge, have investigated the response of the cells to the three-dimensional environment provided by the diverse types of scaffold.

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Three-dimensional culture of human disc cells within agarose or a collagen sponge: assessment of proteoglycan production

Cited by 123 publications

References 17 publications

Effects of Low Glucose Concentrations on Oxygen Consumption Rates of Intervertebral Disc Cells

Effects of Low Glucose Concentrations on Oxygen Consumption Rates of Intervertebral Disc Cells

Vascular tissue engineering by computer-aided laser micromachining

Three-dimensional cultures of osteogenic and chondrogenic cells: A tissue engineering approach to mimic bone and cartilage in vitro

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