Relationship between oxygen consumption rate and cellular activity of mammalian cells cultured in serum-free media

1990 ◽  
Vol 36 (7) ◽  
pp. 759-762 ◽  
Author(s):  
Koji Yamada ◽  
Shigeki Furushou ◽  
Takuya Sugahara ◽  
Sanetaka Shirahata ◽  
Hiroki Murakami
Keyword(s):  
Oxygen Consumption ◽  
Oxygen Consumption Rate ◽  
Mammalian Cells ◽  
Consumption Rate ◽  
Cellular Activity ◽  
Serum Free ◽  
Free Media ◽  
Cells Cultured

scholarly journals 129 RESPIRATION ACTIVITY OF BOVINE EMBRYOS CULTURED IN SERUM-FREE AND SERUM-CONTAINING MEDIA

2005 ◽  
Vol 17 (2) ◽  
pp. 215 ◽  
Author(s):  
H. Abe ◽  
H. Shiku ◽  
S. Aoyagi ◽  
T. Matsue ◽  
H. Hoshi
Keyword(s):  
Oxygen Consumption ◽  
Oxygen Consumption Rate ◽  
Embryo Quality ◽  
Consumption Rate ◽  
Developmental Stages ◽  
Culture Conditions ◽  
Bovine Embryos ◽  
Respiration Activity ◽  
Serum Free ◽  
Consumption Rates

Oxygen consumption is a ubiquitous parameter which can provide valuable information about metabolic mechanisms and embryo quality. Recently, we succeeded in non-invasively and quantitatively determining oxygen consumption of individual bovine embryos by the scanning electrochemical microscopy (SECM). The aim of this study was to assess by SECM the oxygen consumption of individual bovine embryos at different developmental stages cultured in serum-free and serum-supplemented media. Bovine oocytes were matured in IVMD101 medium [Research Institute for the Functional Peptides (IFP), Shimojo, Yamagata, Japan] and inseminated in BO-based medium. For serum-free culture, inseminated ooocytes were cultured to the blastocyst stage in IVD101 medium in an atmosphere of a low oxygen condition (5% CO2/5% O2/90% N2) at 38.5°C. For serum-supplemented culture, inseminated oocytes were cultured in HPM199 medium (IFP) supplemented with 5% calf serum (HPM199 + CS) in the presence of bovine cumulus/granulosa cells in a humidified atmosphere of 5% CO2 in air. Oxygen consumption by individual bovine embryos was non-invasively quantified by the SECM measuring system. Some embryos were prepared for transmission electron microscopy. The oxygen consumption rates are presented in the table. Oxygen consumption rates (F) of the single embryos were low from 2-cell to 8-cell stages (0.45–0.52 × 10−14 mol s−1). In serum-free culture, an increase in oxygen consumption rate was found at the morula (1.03 × 10−14 mol s−1) stage, and blastocysts showed an even higher oxygen consumption rate (1.86 × 10−14 mol s−1). On the other hand, the oxygen consumption of morulae and blastocysts produced in serum-supplemented medium was lower than that of embryos cultured in serum-free medium. Electron microscopic study demonstrated that many of the mitochondria of morulae and blastocycts cultured in HPM199 + CS medium were an immature form, indicating a correlation between respiration activity and development of mitochondria. These results suggest that the culture conditions affect the respiration activity of bovine embryos. The SECM procedures may have a wide application for judging embryo quality and culture conditions for embryos. Table 1. Oxygen consumption rates (F × 10−14 mol s−1) of the bovine embryos at various developmental stages

scholarly journals Acyl-CoA Synthetase Inhibition Protects Clonal Pancreatic Beta-cell from Effects of Chronic Excess Nutrients

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 652-652
Author(s):  
Yuhan Qiu ◽  
Barbara Corkey ◽  
Jude Deeney ◽  
Catherine Li
Keyword(s):  
Oxygen Consumption ◽  
Intracellular Calcium ◽  
Beta Cells ◽  
Oxygen Consumption Rate ◽  
Lipid Droplets ◽  
Consumption Rate ◽  
Insulin Content ◽  
Intracellular Lipid ◽  
Funding Sources ◽  
Cells Cultured

Abstract Objectives A single nucleotide polymorphism (SNP) in the transcription factor 7 like 2 (TCF7L2) gene is strongly associated with Type 2 Diabetes (T2D), and deletion of this SNP has been shown to reduce long chain acyl-CoA synthetase 5 (ACSL5) mRNA level. Previous research in our lab has shown chronic exposure to excess nutrients (glucose and fatty acid (FA)) increases lipid droplets in beta-cells and causes basal insulin hypersecretion, left-shifted glucose-stimulated insulin secretion (GSIS), blunted maximal GSIS and reduced insulin content. We tested the efficacy of a known phenylpyrazole-carboxamide compound (herein named ADIPO C) to reduce accumulation of intracellular lipid droplets and reverse the left-shift of GSIS in beta-cells. Methods INS-1832/13 cells were cultured in RPMI media containing 10% FBS (source of FA) and either 4 or 11 mM glucose. Insulin was measured by HTRF assay (CisBio). Intracellular lipid was detected by fluorescence microscopy using Nile red. Fluorescent Bodipy-FA was used as a surrogate FA in both ACS activity assays and lipid incorporation into cells. Oxygen consumption rate was measured using the Seahorse ion flux analyzer and intracellular calcium was measured in fura 2 loaded cells mounted on an Olympus confocal microscope. Results Adipo C (10–25 µM) acutely inhibited acyl-CoA synthetase activity by up to 40% in cell homogenates and similarly reduced FA incorporation into neutral lipids in INS-1 (832/13) cells measured using thin layer chromatography. Longer exposure (72 hrs) to ADIPO C significantly decreased intracellular lipid droplets, right-shifted GSIS and increased insulin content in cells cultured in excess nutrients. Cells cultured in excess nutrients exhibited both increased basal oxygen consumption rate and intracellular calcium oscillations, which were both reduced with 72 hrs ADIPO C incubation. Conclusions Based on these results, we conclude that Adipo C has a protective effect on beta-cells exposed to a glucolipotoxic environment and thus may prove to have therapeutic potential in the prevention/treatment of T2D. Funding Sources No funding sources to report.

Microbial efficiency in a meromictic reservoir

2008 ◽  
Vol 37 (2) ◽  
Author(s):  
Grażyna Mazurkiewicz-Boroń ◽  
Teresa Bednarz ◽  
Elżbieta Wilk-Woźniak
Keyword(s):  
Carbon Dioxide ◽  
Oxygen Consumption ◽  
Release Rate ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
Metabolic Efficiency ◽  
Sulphur Compounds ◽  
Carbon Dioxide Release ◽  
Microbial Efficiency ◽  
Ion Contents

Microbial efficiency in a meromictic reservoirIndices of microbial efficiency (expressed as oxygen consumption and carbon dioxide release) were determined in the water column of the meromictic Piaseczno Reservoir (in an opencast sulphur mine), which is rich in sulphur compounds. Phytoplankton abundances were low in both the mixolimnion (up to 15 m depth) and monimolimnion (below 15 m depth). In summer and winter, carbon dioxide release was 3-fold and 5-fold higher, respectively, in the monimolimnion than in the mixolimnion. Laboratory enrichments of the sulphur substrate of the water resulted in a decrease in oxygen consumption rate of by about 42% in mixolimnion samples, and in the carbon dioxide release rate by about 69% in monimolimnion samples. Water temperature, pH and bivalent ion contents were of major importance in shaping the microbial metabolic efficiency in the mixolimnion, whilst in the monimolimnion these relationships were not evident.

scholarly journals Correction: Paszkiewicz et al. A Peripheral CB1R Antagonist Increases Lipolysis, Oxygen Consumption Rate, and Markers of Beiging in 3T3-L1 Adipocytes Similar to RIM, Suggesting That Central Effects Can Be Avoided. Int. J. Mol. Sci. 2020, 21, 6639

2021 ◽  
Vol 22 (9) ◽  
pp. 4366
Author(s):  
Rebecca L. Paszkiewicz ◽  
Richard N. Bergman ◽  
Roberta S. Santos ◽  
Aaron P. Frank ◽  
Orison O. Woolcott ◽  
...  
Keyword(s):  
Oxygen Consumption ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
Central Effects

The authors wish to make the following corrections to this paper [...]

scholarly journals Necrostatin-1 Supplementation to Islet Tissue Culture Enhances the In-Vitro Development and Graft Function of Young Porcine Islets

2021 ◽  
Vol 22 (16) ◽  
pp. 8367
Author(s):  
Hien Lau ◽  
Shiri Li ◽  
Nicole Corrales ◽  
Samuel Rodriguez ◽  
Mohammadreza Mohammadi ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Oxygen Consumption ◽  
Insulin Secretion ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
In Vitro Development ◽  
Porcine Islets ◽  
Vitro Development

Pre-weaned porcine islets (PPIs) represent an unlimited source for islet transplantation but are functionally immature. We previously showed that necrostatin-1 (Nec-1) immediately after islet isolation enhanced the in vitro development of PPIs. Here, we examined the impact of Nec-1 on the in vivo function of PPIs after transplantation in diabetic mice. PPIs were isolated from pancreata of 8–15-day-old, pre-weaned pigs and cultured in media alone, or supplemented with Nec-1 (100 µM) on day 0 or on day 3 of culture (n = 5 for each group). On day 7, islet recovery, viability, oxygen consumption rate, insulin content, cellular composition, insulin secretion capacity, and transplant outcomes were evaluated. While islet viability and oxygen consumption rate remained high throughout 7-day tissue culture, Nec-1 supplementation on day 3 significantly improved islet recovery, insulin content, endocrine composition, GLUT2 expression, differentiation potential, proliferation capacity of endocrine cells, and insulin secretion. Adding Nec-1 on day 3 of tissue culture enhanced the islet recovery, proportion of delta cells, beta-cell differentiation and proliferation, and stimulation index. In vivo, this leads to shorter times to normoglycemia, better glycemic control, and higher circulating insulin. Our findings identify the novel time-dependent effects of Nec-1 supplementation on porcine islet quantity and quality prior to transplantation.

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