scholarly journals Effect of fetal bovine serum on in vitro maturation and fertilization of bovine oocytes

2020 ◽  
Vol 8 (5) ◽  
pp. 2187-2191
Author(s):  
Dipannita Baishya ◽  
Arundhati Bora ◽  
J Goswami ◽  
Aunbha Baruah ◽  
DJ Dutta ◽  
...  
Keyword(s):  
Bovine Serum ◽  
In Vitro Maturation ◽  
Fetal Bovine Serum ◽  
Fetal Bovine ◽  
Bovine Oocytes

scholarly journals Fetal bovine serum is associated with polar body degeneration after in vitro maturation of bovine oocytes

2018 ◽  
Vol 68 (3) ◽  
pp. 279
Author(s):  
B. MACÍAS-GARCÍA ◽  
S. MACEDO ◽  
A. ROCHA ◽  
L. GONZÁLEZ-FERNÁNDEZ
Keyword(s):  
Bovine Serum ◽  
Culture Medium ◽  
Fetal Bovine Serum ◽  
Polar Body ◽  
Chromatin Conformation ◽  
Prolonged Incubation ◽  
Vitro Fertilization ◽  
Fetal Bovine ◽  
Bovine Oocytes

In vitro fertilization (IVF) in cattle is commonly used worldwide. Although extensive research has been conducted using different additives in the different IVF steps, little is known regarding how protein type may affect bovine oocytes during the fertilization period. In addition, unlike Tissue Culture Medium 199 (TCM), fertilization medium may induce oocytes’ chromatin degeneration during prolonged incubation in the horse (Modified Whitten’s medium). Thus, in the present work TCM-199 supplemented with either 7 mg/ml of Bovine Serum Albumin (TCM+BSA) or 10% Fetal Bovine Serum (v/v; TCM+FBS) was used. Bovine oocytes were matured in vitro and placed in the previously mentioned media for further 18 hours, in the absence of added sperm (sham fertilization) and their chromatin conformation was evaluated. After IVM, 78.9% of the initial oocytes had reached the MII stage. After sham fertilization, 58.6% of the oocytes in TCM+BSA while just 28.3% in TCM+FBS maintained the MII chromatin conformation (p < 0.05). Subsequent experiments run using PB extruded oocytes and incubated in TCM+BSA and TCM+FBS during sham fertilization, demonstrated that FBS was consistently associated with polar body dissolution or degeneration.

Replacement of fetal bovine serum and FSH with buffalo follicular fluid in in vitro maturation of buffalo oocytes

1996 ◽  
Vol 45 (1) ◽  
pp. 245 ◽  
Author(s):  
S.K. Das ◽  
M.S. Chauhan ◽  
P. Palta ◽  
P.K. Katiyar ◽  
M.L. Madan
Keyword(s):  
Follicular Fluid ◽  
Bovine Serum ◽  
In Vitro Maturation ◽  
Fetal Bovine Serum ◽  
Fetal Bovine

scholarly journals Influence of bovine serum albumin and fetal bovine serum supplementation during in vitro maturation on lipid and mitochondrial behaviour in oocytes and lipid accumulation in bovine embryos

2016 ◽  
Vol 28 (11) ◽  
pp. 1721 ◽  
Author(s):  
Maite del Collado ◽  
Naiara Z. Saraiva ◽  
Flavia L. Lopes ◽  
Roberta C. Gaspar ◽  
Luciana C. Padilha ◽  
...  
Keyword(s):  
Bovine Serum Albumin ◽  
Lipid Content ◽  
Lipid Accumulation ◽  
Bovine Serum ◽  
In Vitro Maturation ◽  
Fetal Bovine Serum ◽  
Bovine Embryos ◽  
Fetal Bovine

Proper oocyte maturation is crucial for subsequent embryo development; however, oocyte mitochondrial and lipid-droplet behaviour are still poorly understood. Although excessive lipid accumulation during in vitro production (IVP) of bovine embryos has been linked with impaired cryotolerance, lipid oxidation is essential for adequate energy supply. Fetal bovine serum (FBS) and bovine serum albumin (BSA) are supplements used during IVP, containing high and low lipid content, respectively. This study aimed to understand how these supplements influence oocyte mitochondrial and lipid behaviour during in vitro maturation (IVM) in comparison to in vivo maturation, as well as their influence on development rates and embryo lipid accumulation during IVP. We demonstrate that only in vivo-matured oocytes maintained correlation between lipid content and active mitochondria. IVM media containing FBS increased total lipid content 18-fold and resulted in higher lipid accumulation in oocytes when compared with media with BSA. IVM using a lower FBS concentration combined with BSA resulted in satisfactory maturation and embryo development and also reduced lipid accumulation in blastocysts. In conclusion, IVM causes changes in mitochondrial and lipid dynamics, which may have negative effects on oocyte development rates and embryo lipid accumulation. Moreover, decreasing FBS concentrations during IVM may reduce embryo lipid accumulation without affecting production rates.

43 MASS VITRIFICATION OF GERMINAL-VESICLE STAGE EQUINE OOCYTES

2016 ◽  
Vol 28 (2) ◽  
pp. 151
Author(s):  
H. S. Canesin ◽  
I. Ortiz ◽  
J. G. Brom-de-Luna ◽  
Y. H. Choi ◽  
K. Hinrichs
Keyword(s):  
Ethylene Glycol ◽  
Dimethyl Sulfoxide ◽  
Bovine Serum ◽  
In Vitro Maturation ◽  
Fetal Bovine Serum ◽  
Control Group ◽  
Cleavage Rate ◽  
Fetal Bovine ◽  
Vitrification Solution

Oocyte cryopreservation has the potential to preserve female genetics. In addition, equine oocytes are not readily available in some areas, and vitrification could be used to accumulate oocytes at remote locations to provide material for research. To preserve large numbers of oocytes, a method for rapid vitrification of multiple oocytes is needed. First, we determined whether immature equine oocytes could be held overnight before vitrification, and we tested the use of a mesh+capillary-action media-removal vitrification platform. Oocytes were collected via ultrasound-guided transvaginal follicle aspiration and randomly allotted to either immediate vitrification or overnight holding (24 to 27 h in 40% M199-Earle’s salts, 40% M199-Hanks’ salts, 20% fetal bovine serum, and 0.3 mM pyruvate) then vitrification. Oocytes were vitrified using different times (1 or 4 min) in vitrification solution and first warming solution: 1v1w, 1v4w, 4v1w, and 4v4w. The base solution was MH (80% M199-Hanks’ salts and 20% fetal bovine serum). Cryoprotectant concentration (vol/vol) was increased in 3 steps until reaching 7.5% dimethyl sulfoxide and 7.5% ethylene glycol. The oocytes were then held in vitrification solution (MH with 15% dimethyl sulfoxide, 15% ethylene glycol, and 0.5 M sucrose) for either 1 or 4 min, according to treatment, and 3 to 10 oocytes were transferred to a 75-μm sterile stainless steel mesh. The mesh was placed on sterile paper to absorb excess medium, then plunged in LN. The oocytes were warmed in MH solution with 1.25 M sucrose for either 1 or 4 min, then placed in 0.62 M and 0.31 M sucrose solutions for 5 min each and undetermined time in MH. After warming, oocytes were cultured for maturation (in vitro maturation) in M199-Earle’s salts, 5 mU mL–1 FSH, and 10% fetal bovine serum. After 30 to 36 h, the oocytes were denuded and stained with Hoechst 33258. Data were analysed by Fisher’s exact test. There were no significant differences (P > 0.05) in rates of meiotic resumption among timing treatments (35, 24, 26, and 39% for 1v1w, 1v4w, 4v1w, and 4v4w, respectively), nor between immediately vitrified (17/55, 31%) and overnight held-vitrified groups (18/56, 32%). In the second experiment, all oocytes were held overnight. They were vitrified and warmed using only the 1v1w and 4v4w schedules, then subjected to in vitro maturation, intracytoplasmic sperm injection, and embryo culture. The MII rate of the control group (27/37, 73%) was higher (P < 0.05) than that for 1v1w (12/33, 36%) or 4v4w treatments (10/35, 29%). The cleavage rate for control (25/27, 93%) was higher than that for 1v1w (5/9, 56%) but not than that for 4v4w (6/9, 67%). Blastocyst rates were 19% (5/27), 11% (1/9), and 0% (0/9) for control, 1v1w, and 4v4w, respectively (P > 0.05). These results indicate that blastocysts may be produced from equine immature oocytes vitrified en masse; however, both the maturation and blastocyst production rates were relatively low. Additional studies are required to improve the efficiency of this technique. This work was supported by the Clinical Equine ICSI Program, Texas A&M University.

FetalClone™ versus fetal bovine serum for in vitro maturation, fertilization and culture

1992 ◽  
Vol 37 (1) ◽  
pp. 212
Author(s):  
M.R. Flood ◽  
D. Kern ◽  
R. Spendlove ◽  
T.D. Bunch
Keyword(s):  
Bovine Serum ◽  
In Vitro Maturation ◽  
Fetal Bovine Serum ◽  
Fetal Bovine

117 EFFECT OF STEPWISE DILUTION ON THE VIABILITY OF FROZEN - THAWED BOVINE OOCYTES MATURED IN VITRO

2007 ◽  
Vol 19 (1) ◽  
pp. 176 ◽  
Author(s):  
A. Hamawaki ◽  
S. Hamano ◽  
M. Yoshikawa ◽  
K. Matsukawa
Keyword(s):  
Bovine Serum ◽  
Room Temperature ◽  
Fetal Bovine Serum ◽  
Calf Serum ◽  
Fertilization Rate ◽  
Single Step ◽  
Normal Morphology ◽  
Fetal Bovine ◽  
Bovine Oocytes

The purpose of this study was to evaluate the effect of stepwise dilution on the viability of frozen–thawed bovine oocytes matured in vitro. Oocytes matured in vitro were denuded and equilibrated in modified TCM-199 (m199: 11 mmol L-1 HEPES, 9 mmol L-1 Na-HEPES, 5 mmol L-1 NaHCO3, 20% (v/v) calf serum) supplemented with 10% (v/v) glycerol for 15 min at room temperature (RT). Then they were exposed to m199 with 10% glycerol and 0.25 mol L-1 sucrose and loaded into 0.25-mL plastic straws. The straws were sealed and seeded at -6�C, cooled at the rate of 0.33�C min-1 to -25�C, and plunged into LN2. For thawing, the straws were first held in air at RT for 10 s, followed by immersion in 30�C water for 10 s. In the first experiment, frozen-thawed oocytes were subjected to cryoprotectants in 5 different manners of dilution. In the non-step dilution, the oocytes (n = 60) were put into m199 for 5 min. In the single-step dilution, the oocytes (n = 37) were transferred to 0.25 mol L-1 sucrose in m199 for 5 min. In the two-step dilution, the oocytes (n = 56) were transferred to 0.5 and then 0.25 mol L-1 sucrose in m199 for 5 and 5 min, respectively. In the three-step dilution, the oocytes (n = 57) were transferred to 0.75, 0.5, and 0.25 mol L-1 sucrose in m199 for 1, 5, and 5 min, respectively. In the four-step dilution, the oocytes (n = 52) were transferred to 1.0, 0.75, 0.5, and 0.25 mol L-1 sucrose in m199 for 1, 1, 5, and 5 min, respectively. After dilution, all of the oocytes were washed twice in TCM-199 supplemented with 5% fetal bovine serum for 5 min and cultured for 1 h to assess the morphology. The rate of morphological normal oocytes in the four-step dilution (94.2%) was significantly (P &lt; 0.05) higher than that in other groups (non-, single-, two-, and three-step dilution: 61.7%, 73.0%, 78.6%, and 77.2%). In the second experiment, non-frozen (control, n = 170) and frozen–thawed oocytes (n = 145) with four-step dilution were fertilized and cultured in vitro (Kuwayama 1992 J. Reprod. Fert. 96, 187–193). To assess fertilization, some of the oocytes were fixed at 10 h after insemination. Cleavage and blastocyst rates were determined on Day 2 and Day 8 after fertilization (Day 0), respectively. There was no difference (P &gt; 0.05) between control and frozen–thawed oocytes in the fertilization rate (88.0% vs. 93.1%). Some of the frozen–thawed oocytes cleaved and developed to blastocysts (44.0% and 11.2%), although the rates were significantly (P &lt; 0.01) lower than those in control (71.7% and 35.0%). These results indicate that stepwise dilution of frozen–thawed oocytes improves the recovery of oocytes with normal morphology, and that the oocytes maintain the abilities to be fertilized and develop to blastocysts.

181 RESVERATROL DURING IN VITRO MATURATION IMPROVES THE QUALITY OF BOVINE OOCYTE AND ENHANCES EMBRYONIC DEVELOPMENT IN VITRO

2017 ◽  
Vol 29 (1) ◽  
pp. 199 ◽  
Author(s):  
V. Torres ◽  
L. Muñoz ◽  
R. Urrego ◽  
J. J. Echeverry ◽  
A. Lopez
Keyword(s):  
Fatty Acid ◽  
Embryonic Development ◽  
Bovine Serum ◽  
In Vitro Maturation ◽  
Fetal Bovine Serum ◽  
Fetal Bovine ◽  
Development In Vitro ◽  
And Control

It is known that reactive oxygen species (ROS) are accumulated within the oocyte during in vitro maturation (IVM) and have been related to poor quality and decreased embryo development in vitro. The use of antioxidants in culture media is an alternative to overcome oxidative stress damage in the oocyte. Resveratrol (3,5,4′-trihydroxy-trans-stilbene) is a phenol produced naturally by several plants and has shown protection against oxidative damage in numerous cell types. Two different experiments were performed to evaluate the effect of resveratrol on the quality of bovine oocytes matured in vitro assessed by levels of ROS and intracellular glutathione (GSH) as well as in vitro embryo development rates. Experiment 1 used different concentrations of resveratrol [0 (Control), 1 (R1), 10 (R10), 20 (R20), and 40 (R40) μM] were used to supplement IVM media. Ovaries were collected from Bos indicus cows at a local abattoir and cumulus-oocyte complexes were matured in vitro for 24 h in TCM199 with 6 mg mL−1 of fatty acid-free BSA, 5% fetal bovine serum, 0.2 mM Na-pyruvate, 50 μg mL−1 of gentamicin, 0.5 μg mL−1 of FSH, and 0.5 µg mL−1 of LH at 38°C in 5% CO2 and 90% humidity. The ROS were evaluated by 2′,7′-dichlorodihydrofluorescein diacetate staining (n = 301) and intracellular GSH levels were determined by Cell Tracker Blue fluorescent stain (n = 310). Denuded oocytes were observed under an epifluorescence microscope. Fluorescence intensities of oocytes were analysed by ImageJ software (Version 1.49v, National Institutes of Health, Bethesda, MD, USA) and normalized to control oocytes. Experiment 2 used cumulus-oocyte complexes (n = 674) collected and matured in vitro under the same conditions described for Exp. 1. In vitro fertilization was performed for 18 h at 38°C in 5% CO2 in Tyrode’s medium with 25 mM bicarbonate, 22 mM Na-lactate, 1 mM Na-pyruvate, and 6 mg mL−1 of fatty acid-free BSA. Additionally 10 µg mL−1 of heparin and 20 μM d-penicillamine, 10 μM hypotaurine, and 1 μM epinephrine were added. The presumptive zygotes were cultured in vitro in SOFaa medium with 5% fetal bovine serum, at 38°C, in 5% CO2, 5% O2, and 90% humidity until Day 7, when embryonic development was assessed. Data were analysed by ANOVA followed by Fisher´s multiple range test using Statgraphics Centurion XVI (Version 16.2.04, Statpoint Technologies Inc., Warrentown, VA). Data are presented as percentage mean ± standard error of the mean (P < 0.05). All concentrations of resveratrol in treated oocytes showed reduced intracellular levels of ROS compared to control (R1: 0.66 ± 0.04, R10: 0.55 ± 0.04, R20: 0.62 ± 0.04, R40: 0.64 ± 0.04, and Control: 1 ± 0.04 pixel/oocyte; P < 0.01). Intracellular levels of GSH were significantly higher for R1 (1.4 ± 0.06; P < 0.01) and R10 (1.3 ± 0.06; P < 0.01) compared with the control. On the other hand, R10 showed a significantly higher blastocyst rate (51% ± 3) compared with R1 (39% ± 4), R20 (39% ± 3), R40 (33% ± 3), and control (38% ± 4). Treatments R1, R20, and R40 showed no significant differences compared to control. These results indicate that resveratrol at 10 μM during IVM improves maturation conditions by decreasing ROS level, increasing intracellular GSH, and improving embryonic developmental competence.

Successful piglet production by IVF of oocytes matured in vitro using NCSU-37 supplemented with fetal bovine serum

2006 ◽  
Vol 65 (2) ◽  
pp. 374-386 ◽  
Author(s):  
Misae Suzuki ◽  
Koji Misumi ◽  
Manabu Ozawa ◽  
Junko Noguchi ◽  
Hiroyuki Kaneko ◽  
...  
Keyword(s):  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Fetal Bovine

scholarly journals Stimulatory Role of Glycated Fetal Bovine Serum Along with Iron on In Vitro Production of Insulin by Differentiated Mouse Embryonic Stem Cells

2011 ◽  
Vol 57 (4) ◽  
pp. 356-361
Author(s):  
Ikuo Nishigaki ◽  
Gowri Rangasamy Gunassekaran ◽  
Panjan Nagappan Venkatesan ◽  
Mandupal Chaco Sabu ◽  
Sabu Priya ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Embryonic Stem ◽  
In Vitro Production ◽  
Fetal Bovine ◽  
Vitro Production

scholarly journals Fetal Bovine Serum-Derived Extracellular Vesicles Persist within Vesicle-Depleted Culture Media

2018 ◽  
Vol 19 (11) ◽  
pp. 3538 ◽  
Author(s):  
Brandon Lehrich ◽  
Yaxuan Liang ◽  
Pooya Khosravi ◽  
Howard Federoff ◽  
Massimo Fiandaca
Keyword(s):  
Cell Growth ◽  
Extracellular Vesicles ◽  
Bovine Serum ◽  
Culture Media ◽  
Fetal Bovine Serum ◽  
Cellular Growth ◽  
Primary Astrocyte ◽  
Fetal Bovine ◽  
Cell Growth And Viability

It is known that culture media (CM) promotes cellular growth, adhesion, and protects explanted primary brain cells from in vitro stresses. The fetal bovine serum (FBS) supplement used in most CM, however, contains significant quantities of extracellular vesicles (EVs) that confound quantitative and qualitative analyses from the EVs produced by the cultured cells. We quantitatively tested the ability of common FBS EV-depletion protocols to remove exogenous EVs from FBS-supplemented CM and evaluated the influence such methods have on primary astrocyte culture growth and viability. We assessed two methodologies utilized for FBS EV removal prior to adding to CM: (1) an 18-h ultracentrifugation (UC); and (2) a commercial EV-depleted FBS (Exo-FBS™). Our analysis demonstrated that Exo-FBS™ CM provided the largest depletion (75%) of total FBS EVs, while still providing 6.92 × 109 ± 1.39 × 108 EVs/mL. In addition, both UC and Exo-FBS™ CM resulted in poor primary astrocyte cell growth and viability in culture. The two common FBS EV-depletion methods investigated, therefore, not only contaminate in vitro primary cell-derived EV analyses, but also provide a suboptimal environment for primary astrocyte cell growth and viability. It appears likely that future CM optimization, using a serum-free alternative, might be required to advance analyses of cell-specific EVs isolated in vitro.

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