Influence of glucose and oxygen supply conditions on the oxygenation of multicellular spheroids

Mueller‐Klieser, Wolfgang; Freyer, James P.; Sutherland, Robert M.

doi:10.1038/bjc.1986.58

Cited by 165 publications

(151 citation statements)

References 20 publications

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“…Only when the PO 2 dropped to zero was the change in consumption large enough to require the addition of a new layer to the model. Similar results were found when PO 2 profiles recorded in tumor spheroids were fitted to diffusion models (Mueller-Klieser, 1984,Mueller-Klieser et al, 1986. These profiles were typically measured until the electrode was about half way through the spheroid.…”

Section: Diffusion Modeling Of Po 2 Profilessupporting

confidence: 73%

“…In larger EMT6/Ro spheroids, the central PO 2 was sometimes nonzero, even when necrosis was present and the PO 2 profile was flat in the center (dP/dr=0) (Mueller-Klieser et al, 1986,Mueller-Klieser and Sutherland, 1982,Mueller-Klieser and Sutherland, 1984. This was also true for other spheroids grown from cells of rodent and human origin (Mueller-Klieser, 1987).…”

Section: Diffusion Modeling Of Po 2 Profilesmentioning

confidence: 87%

“…Thus, these cells were exposed to lower oxygen levels during growth and during the measurements. When EMT6/Ro spheroids were grown on 5% O 2 instead of 20% O 2 , the oxygen consumption rate dropped from 3.09 to 1.65 x 10 −4 ml O 2 /ml/sec (Mueller-Klieser et al, 1986). Therefore, growth at lower oxygen levels significantly decreased spheroid oxygen consumption.…”

Section: Oxygen Consumption Of Ocm-1 MCLmentioning

confidence: 94%

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Modeling of oxygen transport across tumor multicellular layers

Braun

2007

Microvascular Research

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Purpose-Tumor oxygen level plays a major role in the response of tumors to different treatments. The purpose of this study was to develop a method of determining oxygen transport properties in a recently developed 3-D model of tumor parenchyma, the multicellular layer (MCL).Methods-OCM-1 human choroidal melanoma cells were grown as 3-D MCL on collagen-coated culture plate inserts. A recessed-cathode oxygen microelectrode was used to measure oxygen tension (PO 2 ) profiles across 8 different MCL from the free surface to the insert membrane. The profiles were fitted to four different one-dimensional diffusion models: 1-, 2-, and 3-region models with uniform oxygen consumption (q) in each region and a modified 3-region model with a central region where q=0 and PO 2 =0.Results-Depending upon the presence of a central region of anoxia, the PO 2 profiles were fitted best by either the two-region model or the modified 3-region model. Consumption of tumor cells near the insert membrane was higher than that of cells close to the free surface (33.1 ± 13.6 x 10 −4 vs. 11.8 ± 6.7 x 10 −4 mm Hg/m 2 , respectively). Conclusions-The model is useful for determining oxygenation and consumption in MCL, especially for cell lines that cannot be grown as spheroids. In the future, this model will permit the study of parameters important in tumor oxygenation in vitro.

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Section: Diffusion Modeling Of Po 2 Profilessupporting

confidence: 73%

Section: Diffusion Modeling Of Po 2 Profilesmentioning

confidence: 87%

Section: Oxygen Consumption Of Ocm-1 MCLmentioning

confidence: 94%

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Modeling of oxygen transport across tumor multicellular layers

Braun

2007

Microvascular Research

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“…1982). The interactions observed between these nutrients suggested that EMT6 cells in spheroids can adapt their metabolic rates substantially to different supply conditions (Mueller-Klieser et al 1986). as well as to the concentration of lactate (Bourrat-Floeck et al 1991).…”

Section: Resultsmentioning

confidence: 99%

Variable presence of hypoxia in M006 human glioma spheroids and in spheroids and xenografts of clonally derived sublines

Franko¹,

Parliament²,

Allalunis-Turner³

et al. 1998

Br J Cancer

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Summary Recently we reported the variable presence of hypoxia adjacent to necrosis in human glioma lines grown as subcutaneous tumours in severe combined immunodeficient (SCID) mice. To assess the basis for this observation. we examined the pattem of oxygenation in M006 and M006XLo glioma spheroids. We found a wide range of binding of [3H] aExduding two spheroids with a mixture of high and low grains ( Figure 3B).completely at each feeding. The number of spheroids w-as reduced at each feeding to stabilize the rate of consumption of nutrients. The first spheroids reached diameters of 0.8-1.2 mm. at x-hich they w-ere eliaible for experiments. w ithin 4-5 eeks. M006XLo sublineIn an effort to denrve a subline that w-as adapted to grow-th at lowambient oxy gen. the M006XLo subline x as established from small M006 spheroids that had been exposed continuously to 0.6%7 oxygen for 13 days. beainning 1 week after initiation in air. The spheroids were disaggregated wxith 0.25%c trypsin. and the cells w ere groxx n in monolayer culture for four passages (5 w eeks). then injected subcutaneously in SCID mice. A tumour x-as disaggregated to yield a new line designated M006XLo. w hich w as propagated in monolayer culture. Cells from the third passage were stored in liquid nitrogen. and each line x-as discarded after 3 months in culture and replaced from frozen stock.Clonally derived sublines (Franko et al. 1987). The desired oxygen level was obtained by a series of partial evacuations. each of which was followed by refilling of the chambers with 95% nitroaenlr5% carbon dioxide. Metabolic actixation of misonidazole during the degrassing procedure w-as minimized by precooling the medium and the chambers to 0°C and maintainingr this temperature during, the 30-to 40-min degassing process. The incubation period was 3 h for the spinner flask. and an additional 30 min for the aluminum chambers. w-hich is the time required for the medium in the dishes to return to 37' in the cabinet used for incubation. The oxygen tension in each chamber was measured at the end of the incubation period. as described previously (Franko et al. 1987). AutoradiographyThe spheroids were fixed in formalin and embedded in wax. and 4-gm serial sections w-ere taken throughout the spheroids. Slides w-ith sections near the centres of the majority of the spheroids were

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“…We postulate that transformed cells may be sensitive to inhibition of glycolytic flux when oxygen and glucose diffusion limitations are imposed by surrounding cells present in such masses. Specifically, oxygen and glucose diffusion from the culture medium and vasculature may be limited by adjacent cells during soft agar growth and tumor growth, respectively, but remain relatively unlimited for cells grown as a monolayer in vitro (Mueller-Klieser et al, 1986). Accordingly, if glycolytic flux is inadequate, then such diffusion limitations may restrict the availability of energy and anabolic precursors, which then may hinder spheroid or tumor growth.…”

Section: Discussionmentioning

confidence: 99%

Ras transformation requires metabolic control by 6-phosphofructo-2-kinase

et al. 2006

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Neoplastic cells transport large amounts of glucose in order to produce anabolic precursors and energy within the inhospitable environment of a tumor. The ras signaling pathway is activated in several cancers and has been found to stimulate glycolytic flux to lactate. Glycolysis is regulated by ras via the activity of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatases (PFK2/FBPase), which modulate the intracellular concentration of the allosteric glycolytic activator, fructose-2,6-bisphosphate (F2,6BP). We report herein that sequential immortalization and ras-transformation of mouse fibroblasts or human bronchial epithelial cells paradoxically decreases the intracellular concentration of F2,6BP. This marked reduction in the intracellular concentration of F2,6BP sensitizes transformed cells to the antimetabolic effects of PFK2/FBPase inhibition. Moreover, despite co-expression of all four mRNA species (PFKFB1-4), heterozygotic genomic deletion of the inducible PFKFB3 gene in rastransformed mouse lung fibroblasts suppresses F2,6BP production, glycolytic flux to lactate, and growth as soft agar colonies or tumors in athymic mice. These data indicate that the PFKFB3 protein product may serve as an essential downstream metabolic mediator of oncogenic ras, and we propose that pharmacologic inhibition of this enzyme should selectively suppress the high rate of glycolysis and growth by cancer cells.

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Influence of glucose and oxygen supply conditions on the oxygenation of multicellular spheroids

Cited by 165 publications

References 20 publications

Modeling of oxygen transport across tumor multicellular layers

Modeling of oxygen transport across tumor multicellular layers

Variable presence of hypoxia in M006 human glioma spheroids and in spheroids and xenografts of clonally derived sublines

Ras transformation requires metabolic control by 6-phosphofructo-2-kinase

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