Total Anti‐oxidant Capacity of Cell Culture Media

Lewińska, Anna; Wnuk, Maciej; Słota, E.; Bartosz, Grzegorz

doi:10.1111/j.1440-1681.2007.04637.x

Cited by 56 publications

(42 citation statements)

References 36 publications

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“…The antioxidant compounds in SOF medium include methionine, hypotaurine, cystine, tyrosine, tryptophan, besides FBS, BSA, pyruvate and citrate (Guérin et al, 2001;Leminska et al, 2007). Possibly, the similarity of the intracellular ROS levels in the embryos cultured under different O 2 tensions and with different antioxidants may be due to the multiplicity of factors present in the culture media.…”

mentioning

confidence: 97%

Effects of gaseous atmosphere and antioxidants on the development and cryotolerance of bovine embryos at different periods ofin vitroculture

Rocha-Frigoni

Leão

Nogueira

et al. 2013

Zygote

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This study examined the effects of antioxidant supplementation and O2 tension on embryo development, cryotolerance and intracellular reactive oxygen species (ROS) levels. The antioxidant supplementation consisted of 0.6 mM cysteine (CYST); 0.6 mM cysteine + 100 μM cysteamine (C+C); 100 IU catalase (CAT) or 100 μM β-mercaptoethanol (β-ME) for 3 or 7 days of in vitro culture (IVC). Two O2 tensions (20% O2 [5% CO2 in air] or 7% O2, 5% CO2 and 88% N2 [gaseous mixture]) were examined. After 7 days of antioxidant supplementation, the blastocyst frequencies were adversely affected (P < 0.05) by CYST (11.2%) and C+C (1.44%), as well as by low O2 tension (17.2% and 11.11% for 20% and 7% O2, respectively) compared with the control (26.6%). The blastocyst re-expansion rates were not affected (P > 0.05) by the treatments (range, 66-100%). After 3 days of antioxidant supplementation, the blastocyst frequencies were not affected (P > 0.05) by any of the antioxidants (range, 43.6-48.5%), but they were reduced by low O2 tension (P < 0.05) (52.1% and 38.4% for 20% and 7% O2, respectively). The intracellular ROS levels, demonstrated as arbitrary fluorescence units, were not affected (P > 0.05) by antioxidant treatment (range, 0.78 to 0.95) or by O2 tension (0.86 and 0.88 for 20% and 7% O2, respectively). The re-expansion rates were not affected (P > 0.05) by any of the treatments (range, 63.6-93.3%). In conclusion, intracellular antioxidant supplementation and low O2 tension throughout the entire IVC period were deleterious to embryo development. However, antioxidant supplementation up to day 3 of IVC did not affect the blastocyst frequencies or intracellular ROS levels.

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confidence: 97%

Effects of gaseous atmosphere and antioxidants on the development and cryotolerance of bovine embryos at different periods ofin vitroculture

Rocha-Frigoni

Leão

Nogueira

et al. 2013

Zygote

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“…It also shows seasonal and continental variations in the serum composition so that batch-to batch variations are well documented (Van der Valk et al 2010). As its addition is important for cellular antioxidative protection (Lewinska et al 2007), its absence could diminish the total antioxidant capacity of the media, what is particularly important in the case of DMEM due to its high iron and riboflavin content (Table 1). Iron can undergo redox cycling (ferrous-ferric conversion), triggering H 2 O 2 generation, depending on the balance between oxidizing and reducing agents in the media, whereas riboflavin may also induce the formation of ROS on its own way (see above).…”

Section: Resultsmentioning

confidence: 97%

“…FBS is a natural complex mixture of different factors and contains a large number of components like growth inducers, proteins, vitamins, trace elements, hormones, etc., essential for the growth and maintenance of cells (Van der Valk et al 2010). FBS addition is also important for cellular antioxidative protection (Lewinska et al 2007). In its absence the total antioxidant capacity of cell culture media can be diminished.…”

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confidence: 99%

As2O3 oxidation by vitamin C: cell culture studies

et al. 2011

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The ability of As(2)O(3) to induce apoptosis in various malignant cell lines has made it a potential treatment agent for several malignancies. In this study the chemical stability of As(2)O(3) (As(III)) in cell-free growth media with various compositions was studied (MEM with different amount of amino acids and DMEM). Special attention was given to evaluate the influence of serum (FBS; fetal bovine serum) absence and vitamin C addition on the oxidation of As(III) to As(V) in cell-free growth media. FBS is an important source of antioxidants and vitamin C (ascorbic acid) is acting as a prooxidant in millimolar concentrations. Media were incubated with As(III) (0.6, 2 and 7 μmol l(-1)) up to 72 h. Experiments were performed at 37°C in light or/and in the dark, with or without added serum (10%) or vitamin C (1.4, 0.14 mM). Metabolites were followed with high-performance liquid chromatography directly coupled to a hydride generation-atomic fluorescence spectrometry system. After 72 h up to 30% of As(III) was transformed into As(V) in MEMs and up to 35% in DMEM when exposed in dark. Light had no influence on transformations in MEMs, but changed the situation dramatically in DMEM where almost all As(III) was oxidized to As(V) after 72 h when exposed to light. Except for some faster oxidation rate the absence of FBS had little effect on the transformation rate in all media. The most visible impact on As(III) oxidation was observed by addition of vitamin C. Addition of vitamin C (1.4 mM) transformed almost all As(III) to As(V) within 72 h. In lower concentrations (0.14 mM) a pro-oxidative effect was still observed reaching approximately 60% oxidation of As(III) during 72 h. All oxidation processes could be explained by pseudo first order reaction kinetics, yielding reaction rates increasing with initial As(III) concentration and vitamin C concentration whereas the FBS content additionally increased the As(III) oxidation rate in the DMEM (light). The temporal oxidation of As(III) to As(V) in various cell-free growth media necessitates routine checking of the valence state of arsenic during cell culture experiments and the results of biological effects attributed to As(III) should be interpreted with caution. Special attention is needed particularly in cases with vitamin C which was acting pro-oxidatively in all conditions examined.

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“…Plasma Trolox equivalent antioxidant capacity (TEAC) was determined using the method described by Re et al (1999) and modified by Lewinska et al (2007). Briefly, a fresh solution was prepared by dissolving 19.5 mg 2,2 0 -azinobis(3-ethylbenzthiazoline-6-sulphonic acid (ABTS)) and 3.3 mg potassium persulphate in 7 ml of 0.1 mol/l phosphate buffer, pH 7.4.…”

Section: Determination Of Trolox Equivalent Antioxidant Capacity In Pmentioning

confidence: 99%

Melatonin deprival modifies follicular and corpus luteal growth dynamics in a sheep model

Manca¹,

Manunta²,

Spezzigu³

et al. 2014

Reproduction

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This study assessed the effect of melatonin deprival on ovarian status and function in sheep. Experimental procedures were carried out within two consecutive breeding seasons. Animals were divided into two groups: pinealectomised (nZ6) and sham-operated (nZ6). The completeness of the pineal gland removal was confirmed by the plasma concentration of melatonin. Ovarian status was monitored by ovarian ultrasonography for 1 year to study reproductive seasonality. Follicular and corpus luteal growth dynamics were assessed during an induced oestrous cycle. As the effects of melatonin on the ovary may also be mediated by its antioxidant properties, plasma Trolox equivalent antioxidant capacity (TEAC) was determined monthly for 1 year. Pinealectomy significantly extended the breeding season (310G24.7 vs 217.5G24.7 days in controls; P!0.05). Both pinealectomised and sham-operated ewes showed a well-defined wave-like pattern of follicle dynamics; however, melatonin deficiency caused fewer waves during the oestrous cycle (4.3G0.2 vs 5.2G0.2; P!0.05), because waves were 1 day longer when compared with the controls (7.2G0.3 vs 6.1G0.3; P!0.05). The mean area of the corpora lutea (105.4G5.9 vs 65.4G5.9 mm 2 ; P!0.05) and plasma progesterone levels (7.1G0.7 vs 4.9G0.6 ng/ml; P!0.05) were significantly higher in sham-operated ewes compared with pinealectomised ewes. In addition, TEAC values were significantly lower in pinealectomised ewes compared with control ones. These data suggest that melatonin, besides exerting its well-known role in the synchronisation of seasonal reproductive fluctuations, influences the growth pattern of the follicles and the steroidogenic capacity of the corpus luteum. Free Italian abstractAn Italian translation of this abstract is freely available at

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Total Anti‐oxidant Capacity of Cell Culture Media

Cited by 56 publications

References 36 publications

Effects of gaseous atmosphere and antioxidants on the development and cryotolerance of bovine embryos at different periods ofin vitroculture

Effects of gaseous atmosphere and antioxidants on the development and cryotolerance of bovine embryos at different periods ofin vitroculture

As2O3 oxidation by vitamin C: cell culture studies

Melatonin deprival modifies follicular and corpus luteal growth dynamics in a sheep model

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