Determination of Gaseous and Dissolved Oxygen in a Closed Fat Cell Culture System

Böttcher, Hartfried; Engel, Sabine; Fürst, Peter

doi:10.1006/abio.1995.1449

Cited by 8 publications

(9 citation statements)

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“…Decreased fat cell thermogenesis in obese humans H Bo Èttcher and P Fu Èrst derived from human adipocytes in vivo or in agarose gel cultures and suspensions. 19,23,26 Considering the positive correlation of cell size and the rates of oxidative phosphorylation, glycolysis and lipolysis, 5,15,27±30 it is not likely that the observed decrease of thermogenic rate with increasing BMI (Figure 1) is due to an increased fat cell size. 31 Indeed, human obesity is associated with increased fat cell thermogenesis despite a larger cell size.…”

Section: Basal Conditionsmentioning

confidence: 98%

“…From each preparation, up to 20 parallel cultures were established and kept at 37 C. The procedure was described earlier in detail. 19,22,23 Under these culture conditions, basal rates of fat cell thermogenesis, lipolysis, glycolysis and oxidative phosphorylation are constant for more than 72 h, while FFA release declines continuously. 19,23 Microcalorimetry Two identical microcalorimeters of the thermopile heat conduction type (2277-BioActivityMonitor, LKB, Bromma, Sweden and 2277-ThermalActivityMonitor, ThermoMetric, Ja Èrfa Èlla, Sweden) were used.…”

Section: Fat Cell Culturementioning

confidence: 98%

“…These methods were previously described and evaluated in detail. 19,23 Re-esteri®cation rate of FFA (R RE ) was calculated from the lipolytic rate (R GLL ) and the rate of FFA release (R FFA ) according to the formula R RE 3 6 R GLL 7 R FFA . 15 …”

Section: Biochemcial Analysesmentioning

confidence: 99%

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Decreased white fat cell thermogenesis in obese individuals

Böttcher

Fürst

1997

Int J Obes

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OBJECTIVE: To investigate whether white adipocyte thermogenesis and energy metabolism are reduced in obese individuals. SUBJECTS: Eight lean and 15 obese men and women; BMI 19±41. DESIGN: Isolated subcutaneous adipocytes were maintained in agarose gel for 20 h under basal conditions and subsequently for 10 h after stimulation with 1 mM isoprenaline. Direct microcalorimetry was performed continuously over 30 h while biochemical measures were obtained after 0, 20, 25 and 30 h. MEASUREMENTS: Total cellular thermogenesis, oxygen consumption, glycolysis, lipolysis, triglyceride/FFA substrate cycle, adenine nucleotides, DNA content as basis of reference. RESULTS: Under basal and stimulated conditions, thermogenesis (5.6 and 8.6 mW/mgDNA, respectively; P < 0.0001) correlated negatively (P < 0.01 and P < 0.05, respectively) with the BMI and positively with O 2 and glucose consumption, lactate, glycerol, FFA release and FFA re-esteri®cation. Reduced basal lactate production with increased BMI (P < 0.05) indicates a more aerobic adipocyte metabolism in obese individuals. Negative correlation between BMI and stimulated triglyceride/FFA substrate cycle activity (P < 0.01) explains the decreased hormone induced adipocyte heat production in the obese. CONCLUSIONS: The results suggest that a reduction of total body energy expenditure, which is discussed to cause obesity, can be associated with distinct metabolic alterations at a cellular level. However, since the estimated total body fat cell thermogenesis does not exceed 7% of the resting metabolic rate, the observed decrease of adipocyte heat production in the face of augmented BMI can only in part be responsible for the development of obesity.

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Section: Basal Conditionsmentioning

confidence: 98%

Section: Fat Cell Culturementioning

confidence: 98%

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Decreased white fat cell thermogenesis in obese individuals

Böttcher

Fürst

1997

Int J Obes

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“…The only other system that has been used to measure long-term metabolic rates of cell cultures is that of Bottcher et al (1995). This appears to be a promising system.…”

Section: Discussionmentioning

confidence: 97%

“…There is to our knowledge only one report of oxygen consumption measurements in long-term cell culture. This is in human fat cells embedded in agarose medium (Bottcher et al, 1995). This study will be addressed more closely in Discussion.…”

mentioning

confidence: 98%

Method for measuring a comprehensive energy budget in a proliferating cell system over multiple cell cycles

et al. 1997

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Isolated cell systems are now being used very effectively to study a range of important biochemical questions, but their energy metabolism has never been comprehensively investigated. We have developed a system, using J2E cells, which enables us to measure total ATP turnover and the contribution of various fuels and pathways to this total in a dynamic, proliferating preparation. Cells are cultured in 500 ml airtight glass containers which enables (1) the measurement of oxygen consumption, (2) the collection and measurement of 14CO2 production from labelled fuels, and (3) the measurement of metabolite utilization and production. Data on cell numbers are then used to produce a curve of cell number vs. time, the area under which (cell numbers x hour) is used as a base by which all measurements and experiments are compared. To our knowledge this is the first time a comprehensive energy budget has been measured in a proliferating cell system over a period that covers multiple cell cycles.

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Method for determining oxygen consumption rates of static cultures from microplate measurements of pericellular dissolved oxygen concentration

Guarino

Dike

Haq

et al. 2004

Biotech & Bioengineering

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We describe a simple protocol for determining the oxygen consumption of cells in static culture. The protocol is based on a noninvasive oxygen-sensing microplate and a simple mathematical model derived from Fick's Law. The applicability of the model is confirmed by showing the correlation of computed oxygen consumption rate (OCR) values to actual cell densities ascertained by direct cell counting and/or MTT for HL60 and U937 cells cultured in suspension. Correlation between computed OCR and these other indications of cell number was quite good, as long as the cultures were not diffusion-limited for oxygen. The impact of the geometric factors of media depth and well size were confirmed to be consistent with the model. Based on this demonstrated correlation, we also developed a simple, completely noninvasive algorithm for ascertaining the per-cell oxygen utilization rate (OUR), which is the ratio of OCR to cell number, and a fundamental cell characteristic. This is accomplished by correlating the known seed densities to extrapolated determinations of OCR at time zero. Such determinations were performed for numerous cell types, in varying well sizes. Resulting OUR values are consistent with literature values acquired by far more painstaking methods, and ranged from <0.01 fmol.min(-1).cell(-1) for bacteria to 0.1-10 fmol.min(-1).cell(-1) for immortalized mammalian and insect cell lines to >10 fmol.min(-1).cell(-1) for primary hepatocytes. This protocol for determining OCR and OUR is extremely simple and broadly applicable and can afford rapid, informative, and noninvasive insight into the state of the culture.

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Determination of Gaseous and Dissolved Oxygen in a Closed Fat Cell Culture System

Cited by 8 publications

References 0 publications

Decreased white fat cell thermogenesis in obese individuals

Decreased white fat cell thermogenesis in obese individuals

Method for measuring a comprehensive energy budget in a proliferating cell system over multiple cell cycles

Method for determining oxygen consumption rates of static cultures from microplate measurements of pericellular dissolved oxygen concentration

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