Effects of Fractions of Bovine Follicular Fluid and Fetal Bovine Serum as Supplements to Maturation Medium on In Vitro Development of In Vitro Fertilized Bovine Embryos

2011 ◽  
Vol 28 (1) ◽  
pp. 68-74 ◽  
Author(s):  
Kenji Momozawa ◽  
Yoshinori Fukuda
Keyword(s):  
Follicular Fluid ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Bovine Embryos ◽  
In Vitro Development ◽  
Maturation Medium ◽  
Fetal Bovine ◽  
Vitro Development

Effect of charcoal:dextran stripped fetal bovine serum on in vitro development of bovine embryos

2017 ◽  
Vol 17 (4) ◽  
pp. 312-319 ◽  
Author(s):  
Ayman Mesalam ◽  
Rami Kong ◽  
Imran Khan ◽  
MMR Chowdhury ◽  
Byung-Hyun Choi ◽  
...  
Keyword(s):  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Bovine Embryos ◽  
In Vitro Development ◽  
Fetal Bovine ◽  
Vitro Development

184 EFFECT OF CONCENTRATION AND EXPOSURE DURATION OF FETAL BOVINE SERUM ON PARTHENOGENETIC DEVELOPMENT OF PORCINE FOLLICULAR OOCYTES

2007 ◽  
Vol 19 (1) ◽  
pp. 208
Author(s):  
H. J. Kim ◽  
S. R. Cho ◽  
C. Y. Choe ◽  
S. H. Choi ◽  
D. S. Son ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Follicular Fluid ◽  
Bovine Serum ◽  
Exposure Duration ◽  
Cytochalasin B ◽  
Fetal Bovine Serum ◽  
Hatching Rate ◽  
Parthenogenetic Development ◽  
Maturation Medium ◽  
Fetal Bovine

The aim of the present experiment was to examine hatching rate as a testing tool of porcine embryo viability before early-stage embryo transfer, such as zygotes or 2-cell stage embryos. We evaluated the optimal concentrations and exposure durations of fetal bovine serum (FBS) on porcine parthenotes. Ovaries were obtained from prepubertal gilts at a local abattoir and brought to the laboratory in physiological saline with antibiotics at 30–33°C. The ovaries were washed and wiped, and then cumulus–oocytes complexes (COCs) in the follicular fluid were aspirated from surface-visible follicles (2–6 mm in diameter) with a 10-mL syringe fitted with an 18-gauge needle. After being washed 3 times with modified phosphate-buffered saline (DPBS; GIBCO, Grand Island, NY, USA) containing 0.3% BSA, the COCs were suspended in maturation medium, NCSU-23 containing 10% (v/v) porcine follicular fluid, 10 ng mL−1 epidermal growth factor (EFG; Sigma-Aldrich Corp., St Louis, MO, USA), 10 µg mL−1 follicular stimulating hormone (FSH; Sigma), 35 µg mL−1 luteinizing hormone (LH; Sigma), 1 mg mL−1 cysteine (Sigma, USA), 100 IU mL−1 penicillin G, and 100 µg mL−1 streptomycin sulfate (GIBCO). After 24 h, the COCs were transferred to the same medium without hormones. The oocytes matured for 48 h were denuded. The oocytes with a visible polar body were selected and returned to the maturation medium without hormones. After 65 h of maturation, oocytes were exposed to PBS with 7% ethanol (v/v) for 7 min, and then the oocytes were washed and treated in TCM-199 containing 5 µg mL−1 cytochalasin B (Sigma) for 5 h at 38.5°C in an atmosphere of 5% CO2 and 95% air with high humidity. After cytochalasin B treatment, the presumptive parthenotes were cultured in PZM-5 medium (IFP, Japan) and cleavage of the parthenotes was assessed at 72 h of activation. Normally cleaved parthenotes were cultured for 8 days to evaluate their ability to develop to the blastocyst and hatching stages. The FBS was added at Day 4 or 5 with concentrations of 2.5, 5, or 10%. The blastocyst rates ranged from 39.1 to 70% in each treatment. However, the hatching rate was dramatically decreased in the non-addition group. In this experiment, the developmental potential may be estimated before embryo transfer by an in vitro culture system up to the hatching stage. Table 1. Effect of concentration and exposure duration of FBS on parthenogenetic development of porcine follicular oocytes

302 REPLACEMENT OF FETAL BOVINE SERUM WITH KNOCKOUT SERUM REPLACER AND STEM CELLS CONDITIONED MEDIUM DURING IN VITRO CULTURE OF BOVINE EMBRYOS

2010 ◽  
Vol 22 (1) ◽  
pp. 307
Author(s):  
M. M. Souza ◽  
N. Z. Saraiva ◽  
C. S. Oliveira ◽  
T. A. D. Tetzner-Nanzeri ◽  
R. Vantini ◽  
...  
Keyword(s):  
Stem Cells ◽  
In Vitro Culture ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Protein Supplementation ◽  
Control Group ◽  
Bovine Embryos ◽  
Fetal Bovine

The use of fetal bovine serum (FBS) as protein supplementation in IVP of bovine embryos has presented difficulties because it can introduce a number of pathogenic components in culture systems, can be related to the birth of calf with abnormal growth and development, and precludes the establishment of the actual nutritional needs of the embryo, because it contains an unlimited variety of substances. This study evaluated the replacement of the FBS in the medium of in vitro culture (IVC) of bovine embryos, using the knockout serum replacer (KSR) as protein supplementation and culture medium conditioned with stem cells. Therefore, bovine oocytes from ovaries of slaughterhouse were selected and matured in vitro in TCM-199 medium supplemented with 10% FBS (Crypion), 1.0 μg mL-1 FSH (Pluset®, Calier, Barcelona, Spain), 50 μg mL-1 hCG (Profasi®, Serono, Geneva, Switzerland), 1.0 μg mL-1 estradiol (Sigma E-2758, Sigma Chemical, St. Louis, MO, USA), 0.2 mM sodium pyruvate, and 83.4 μg mL-1 amikacin for 24 h. After that, 1144 oocytes were fertilized in IVF-TALP medium containing 6 mg mL-1 of BSA. After 18 to 22 h, the zygotes were cultured in SOF + 5% FBS (group 2); SOF + 5% KSR (group 3); SOF (5% FBS) + 10% SOF (5% FBS) conditioned by stem cells (group 4); or SOF (5% KSR) + 10% SOF (5% KSR) conditioned by stem cells (group 5), in an atmosphere of 5% O2 at 38.5°C for 8 days. A control group outside the controlled atmosphere was added, supplemented with 5% FBS (group 1). The SOF medium supplemented with 5% FBS or KSR was conditioned by stem cells and added to SOF medium for the culture of embryo at a concentration of 10%. The rates of cleavage and production of blastocysts were assessed 48 hours and 7 days after IVF, respectively, and analyzed by chi-square test, with a significance level of 5% in the statistical program Minitab® (release 14.1, Minitab, State College, PA, USA). On the eighth day, the TUNEL test for determination of the percentage of apoptosis and the differential staining technique for determination of inner cell mass (ICM) and trophoblast (TF) were performed. The results were submitted to ANOVA, followed by comparing the means by Tukey’s test using the program GraphPad Prism (GraphPad, San Diego, CA, USA). The treatments did not differ in the production of embryos, being similar to the control group: G1 = 31.75% (74/233), G2 = 35.26% (79/224), G3 = 32.70% (74/226), G4 = 28.76% (63/219), and G5 = 26.85% (65/242). With regard to the assessment of embryonic quality, the treatments showed similar results to the control groups. No differences were observed among groups both in color and ICM/TF ratio (G1 = 0.60, G2 = 0.62, G3 =0.65, G4 = 0.60, and G5 = 0.60). Furthermore, the TUNEL showed no significant difference in the percentage of apoptosis among groups (G1 = 7.10%, G2 = 3.76%, G3 = 5.58%, G4 = 4.50%, and G5 = 4.11%). The data obtained so far indicate that it is possible to produce embryos in vitro by replacing the FBS in the culture, achieving results similar to those obtained with serum. Financial support: FAPESP 2007/58506-6.

Is it possible to alter the embryo lipid accumulation with reduction of fetal bovine serum and use of l-carnitine for in vitro maturation of bubaline oocytes?

Zygote ◽  
2020 ◽  
Vol 28 (2) ◽  
pp. 109-115
Author(s):  
Marivaldo Rodrigues Figueiró ◽  
Joaquim Mansano Garcia ◽  
Marina Ragagnin de Lima ◽  
Maite del Collado ◽  
Naiara Zoccal Saraiva
Keyword(s):  
Lipid Accumulation ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Time Frame ◽  
Success Rates ◽  
High Concentration ◽  
Embryo Production ◽  
Maturation Medium ◽  
Fetal Bovine

SummaryIn vitro embryo production (IVEP) is a procedure that can promote genetic improvement in a short time frame. However, the success rates obtained with this biotechnology in water buffaloes are still inconsistent, and can be associated with the high concentration of lipids in the cytoplasm of oocytes and embryos. The objective of this study was to evaluate the effects of reduced concentration of fetal bovine serum (FBS) and/or use of l-carnitine during in vitro maturation (IVM) on the preimplantation development and lipid accumulation in bubaline embryos. In a first experiment, the lowest concentration of FBS in the IVM medium (0%, 2.5%, 5% or 10%) was determined, and the lowest concentration that maintained good embryo development rates was 5%. In a second experiment, the addition of 5 mM of l-carnitine into the maturation medium was evaluated. The blastocysts produced were submitted to lipid evaluation involving staining followed by observation using optical (Oil Red O) and confocal (BODIPY 493/503) microscopy. No difference was observed between the 5% and 10% FBS groups, which were superior to the 0% and 2.5% groups. Furthermore, the performance of the groups treated with 5% and 10% FBS was better than the groups supplemented with l-carnitine. There was no difference regarding embryo lipid accumulation. The results indicated that it is possible to reduce the FBS concentration to 5% in in vitro maturation medium for production of bubaline embryos, and supplementation with 5 mM l-carnitine does not increase embryo production.

Influence of follicular fluid from different sources on in vitro development of bovine embryos produced in vitro

1997 ◽  
Vol 47 (1) ◽  
pp. 292
Author(s):  
C Larocca ◽  
S Kmaid ◽  
I Lago ◽  
G Roses ◽  
D Fila ◽  
...  
Keyword(s):  
Follicular Fluid ◽  
Bovine Embryos ◽  
In Vitro Development ◽  
Vitro Development ◽  
Different Sources

Distinct effect of fetal bovine serum versus follicular fluid on multipotentiality of human granulosa cells in in vitro condition

Biologicals ◽  
2018 ◽  
Vol 52 ◽  
pp. 44-48 ◽  
Author(s):  
Soudabe Yousefi ◽  
Jafar Soleimanirad ◽  
Kobra Hamdi ◽  
Laya Farzadi ◽  
Aalie Ghasemzadeh ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Follicular Fluid ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Distinct Effect ◽  
Human Granulosa Cells ◽  
Fetal Bovine

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
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