Changes in O2 Consumption of Multicellular Spheroids During Development of Necrosis

Mueller‐Klieser, Wolfgang; Bourrat, B.; Gabbert, Helmut E.; Sutherland, Robert M.

doi:10.1007/978-1-4684-3291-6_78

Cited by 15 publications

(6 citation statements)

References 2 publications

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“…The data presented here suggest that considerable changes in Q may also occur in EMT6 spheroids during growth, but these changes may be distributed over a wider diameter range than in the case of V79 spheroids. This is confirmed by recent observations in EMT6 spheroids cultured under standard growth conditions (Freyer & Sutherland, 1985c;Mueller-Klieser et al, 1986). Since similar changes may be true for all the culture conditions investigated (see Figure 6), changes in Q during spheroid growth may, therefore, not be a result of one specific supply situation but rather a general characteristic of three-dimensional growth of tumour cells.…”

Section: F Discussionsupporting

confidence: 87%

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

Mueller‐Klieser¹,

Freyer²,

Sutherland³

1986

Br J Cancer

165

133

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Summary The interrelationship among external 02 and glucose supply, oxygenation status, oxygen consumption rates and cellular viability in tumour microregions was studied using the multicellular spheroid model. For chronic exposure to various supply conditions multicellular EMT6/Ro spheroids were cultured in stirred media equilibrated either with 20% (v/v) or 5% (v/v) oxygen and containing four different glucose concentrations ranging from 0.8mm to 16.5mM. Spheroids were investigated using histology and 02-sensitive microelectrodes for measuring oxygen tension (PO ) values. A chronic decrease of the glucose concentration in the medium is associated with a substantial reduction in the thickness of the viable rim of cells and with a persistent increase in the cellular respiration rate. In general, both viable rim size and respiration are decreased through restriction of 02 supply during spheroid growth at a given external glucose concentration. The 02 consumption in spheroids appears to decrease with increasing spheroid size under most of the growth conditions investigated. These findings provide evidence for a large capacity of the spheroid cells to chronically adapt their metabolic rates to different supply situations. The experimental data and theoretical considerations indicate that necrosis may develop in the centre of these spheroids due to the lack of 02 and/or glucose under some of the growth conditions, but central necrosis can also occur despite sufficient 02 and glucose supply. Consequently, cellular metabolism and viability in tumour microregions may not be determined by the diffusion limitation of O2 or specific substrates alone, such as glucose, but may be influenced by a complex interaction of factors in the micromilieu the majority of which are still unknown.

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Section: F Discussionsupporting

confidence: 87%

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

Mueller‐Klieser¹,

Freyer²,

Sutherland³

1986

Br J Cancer

165

133

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“…The decreasing oxygen tension in the center of the growing spheroid (Mueller-Klieser et al, 1985) increases lactate production previously shown by MR spectroscopy (Kotitschke et al, 1995). Accordingly, large spheroids made from cell lines express VEGF, mainly in the inner layers of the spheroid .…”

Section: Role Of Vegf In the Spheroid Modelmentioning

confidence: 95%

Vascularization of human glioma spheroids implanted into rat cortex is conferred by two distinct mechanisms

et al. 1999

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Aim of this study was to develop and characterize an applicable in vivo model to investigate angiogenesis of human gliomas. An established glioblastoma spheroid model was used to investigate the neovascularization of a standardized avascular solid tumor mass. Spheroids of two human glioma cell lines were labeled with an in vivo fluorescent dye. Single spheroids were implanted into the cortex of athymic rats. After 1, 3, 7, 14, and 21 days, brain sections containing the spheroid were immunostained for endothelial cells or vascular endothelial growth factor (VEGF). The dye-stained glioma spheroid and the endothelial cells were visualized by confocal microscopy. Two distinct mechanisms of tumor vascularization could be observed. (1) "Classical" angiogenesis with new vessels sprouting from existing host vessels into the spheroid was seen. (2) Individual endothelial cells were found to migrate towards and into the center of the spheroid where they coalesced to form new vessels. This process occurred as early as 24 hr after spheroid implantation. Spheroid vascularization was accompanied by an increase of VEGF expression, which peaked 7 days after implantation and returned to normal patterns by 14-21 days. Besides the "classical" angiogenesis by angiogenic blood vessels, the recruitment of individual endothelial cells seems to be an additional mechanism in early glioma vascularization. Our model proves to be a reliable, reproducible system to study in vivo angiogenesis of human gliomas.

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“…Figure 1) imply a uniformly distributed cellular respiratory activity ( Bourrat‐Floeck et al . 1991 ; Groebe & Mueller‐Klieser 1991; Mueller‐Klieser 1984; Mueller‐Klieser et al . 1985 ), while selective dissociation of the same spheroid types followed by in situ oxygen consumption measurement showed a reduction in oxygen uptake per cell as a function of depth within the spheroid ( Freyer 1994).…”

Section: Spheroid Monoculturesmentioning

confidence: 99%

“…Estimations of oxygen consumption rates from oxygen partial pressure gradients within spheroids (i.e. Figure 1) imply a uniformly distributed cellular respiratory activity (Bourrat-Floeck et al 1991;Groebe & Mueller-Klieser 1991;Mueller-Klieser 1984;Mueller-Klieser et al 1985), while selective dissociation of the same spheroid types followed by in situ oxygen consumption measurement showed a reduction in oxygen uptake per cell as a function of depth within the spheroid (Freyer 1994). Morphometric studies by Bredel-Geissler et al (1992) document a relatively constant number of mitochondria across the viable cell rim of spheroids, yet, without taking into account their functional status.…”

Section: Microenvironmentmentioning

confidence: 99%

Multicellular spheroids: a three‐dimensionalin vitroculture system to study tumour biology

Kunz-Schughart

Kreutz

Knuechel

1998

Int J Experimental Path

305

231

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The growth of tumour cells as three-dimensional multicellular spheroids in vitro has led to important insights in tumour biology, since properties of the in vivo-tumour such as proliferation or nutrient gradients, can be studied under controlled conditions. While this review starts with an update of recent data on spheroid monocultures, especially concerning tumour microenvironment and therapeutic modalities, the main emphasis is put on the spectrum of heterologous cultures which have evolved in previous years. This type of culture includes tumour cell interaction with endothelial, fibroblast or immunocompetent cells. The relation of the spheroid culture model to other types of three-dimensional culture and our critical evaluation and presentation of the technical aspects of growing and analysing spheroids are included in the text. These topics are chosen to help the experimental pathologist design experiments with tumour spheroids and to stimulate discussion.

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Changes in O2 Consumption of Multicellular Spheroids During Development of Necrosis

Cited by 15 publications

References 2 publications

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

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

Vascularization of human glioma spheroids implanted into rat cortex is conferred by two distinct mechanisms

Multicellular spheroids: a three‐dimensionalin vitroculture system to study tumour biology

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