The influence of oxygen on viability and proliferation in cellular spheroids

Carlsson, Jörgen; Stålnacke, Carl-Göran; Acker, H.; M, Haji-Karim; Nilsson, Sverker; Larsson, B.

doi:10.1016/0360-3016(79)90953-2

Cited by 74 publications

(27 citation statements)

References 19 publications

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“…There is evidence from the present data, from data published previously (Carlsson et al, 1979;Mueller-Klieser & Sutherland, 1982a, b;MuellerKlieser et al, 1983) as well as from theoretical considerations (Mueller-Klieser, 1984;Freyer & Sutherland, 1985a, b) that necrosis in spheroids may develop despite sufficient 02 supply. This is true for most of the supply conditions investigated, yet in media with 5.5 mm glucose and a Po2 of 35-40mmHg the central Po2 had dropped to 0mmHg before necrosis arises in the spheroid centre.…”

Section: F Discussionsupporting

confidence: 70%

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

Mueller‐Klieser¹,

Freyer²,

Sutherland³

1986

Br J Cancer

164

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: 70%

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

Mueller‐Klieser¹,

Freyer²,

Sutherland³

1986

Br J Cancer

164

133

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“…Observations of such an effect were initially made decades ago (Warburg, 1956), and experiments with V79 and EMT6 spheroids have shown such an effect (Freyer et al, 1984;Freyer and Sutherland, 1985). Also, necrosis has been observed in human glioma spheroids at relatively high central oxygen levels as determined by microelectrodes (Carlsson et al, 1979(Carlsson et al, , 1983. If down-regulation of oxygen consumption was coordinated at a microregional level, it might provide an explanation for the variable presence of severe hypoxia adjacent to necrosis, as indicated by nitroimidazole binding in the xenografts.…”

Section: Discussionmentioning

confidence: 99%

Anomalous patterns of nitroimidazole binding adjacent to necrosis in human glioma xenografts: possible role of decreased oxygen consumption

Parliament¹,

Franko²,

Allalunis-Turner³

et al. 1997

Br J Cancer

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Summary In contrast to reports of extensive hypoxia in human gliomas in situ measured by P02 histography, non-invasive methods of assessing glioma oxygenation, including nitroimidazole binding, have yielded surprisingly contradictory results. In order to investigate the relationship of necrosis, hypoxia, nitroreductase activity and cellular respiration in human gliomas, subcutaneous models using the human glioma cell lines M059K, M006 and M01 Ob were developed in the murine SCID host. Intracranial growth of the M006 line was achieved in nude rats. The nitroreductive capacity of glioma cell lines was assessed and found to be similar to transplanted tumours previously reported in the literature. This suggests that if substantial numbers of viable hypoxic cells were present in situ in gliomas, then nitroimidazole-binding techniques should be capable of identifying them. Inter-tumour variability in the amount of necrosis was seen. M006 xenografts growing in subcutaneous and intracranial sites revealed extensive necrotic regions histologically, some of which were surrounded by cells labelled heavily for [3H]misonidazole, while other areas were lightly labelled. Similar binding patterns were seen for subcutaneous M059K tumours, while subcutaneous M01 Ob tumours display necroses of which almost all were surrounded by heavily labelled cells. The oxygen consumption rates of tumour cell lines grown in vivo, in which venous P02 concentrations are of the order of 2-5%, were two to sevenfold less than those of the same lines grown as monolayers in vitro under oxygen concentrations of 18%. We postulate that glioma cell lines behave as 'oxygen conformers', in that their rate of oxygen consumption appears to vary with the availability of oxygen. Together with the misonidazole-binding data, the results in this glioma tumour model are consistent with coordinate inhibition or down-regulation of respiration under moderate hypoxia.

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“…This may be due to restricted supply of substrates and inadequate removal of metabolic waste in the attached spheroid (which is suspended in static medium for several hours). Finally, it cannot be excluded that the V-79 strain used by Carlsson et al (1979) had an 02 consumption different from the cell line used in this study. However it can be stated that, despite those differences in measured Po2, necrosis develops in V-79 spheroids in the 2 laboratories under both experimental conditions, where the Po2 is not severely low.…”

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confidence: 92%