162 EFFECT OF MEIOTIC ARREST USING BUTYROLACTONE I AND ROSCOVITINE IN RESISTANCE TO EMBRYO CRYOPRESERVATION

2014 ◽  
Vol 26 (1) ◽  
pp. 195
Author(s):  
R. R. D. Maziero ◽  
D. M. Paschoal ◽  
M. D. Guastali ◽  
M. J. Sudano ◽  
P. N. Guasti ◽  
...  
Keyword(s):  
Breeding Systems ◽  
Embryo Cryopreservation ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Cattle Breeding ◽  
Meiotic Arrest ◽  
Cell Groups ◽  
Level Of Significance ◽  
Vitro Production

In vitro embryo production by maturation, fertilization, and culture has become a valuable assisted reproductive technology in cattle breeding systems. However, even with the notable innovations in this system, the greatest obstacles for the complete success of this biotechnology are the low pregnancy rates after transfer and the greater sensitivity to cryopreservation. Thus, this study evaluated the effect of the addition of a meiotic arrest using butyrolactone I and roscovitine on in vitro production of bovine embryos. Nelore oocytes were matured in TCM-199 with Earle's salt + 10% FCS, FSH, and LH, in 5% CO2 atmosphere. To delay meiosis, the oocytes were maintained for 6 h in medium in presence of butyrolactone I (BL-I) 25 μM + roscovitine (ROS) 6.25 μM. Then, the oocytes (Control n = 405; BL-I + ROS n = 386) were cultured for 18 h in agent-free medium to resume meiosis, completing 24 h of maturation. After 24 h of maturation (Day 0), oocytes were fertilized in human tubal fluid (HTF, Irvine, New Zealand) under the conditions above. Semen was selected through Percoll gradient and the concentration adjusted to 2 × 106 sperm mL–1. The presumed zygotes were culture in 90-μL droplets of SOFaa + 0.6% BSA + 2.5% FCS in 5% CO2 until Day 7, when the blastocyst rate was evaluated. We performed 5 replicates. Data were analysed with ANOVA, followed by Tukey test using the general linear model (PROC GLM) of SAS 9.2 (SAS Institute Inc., Cary, NC, USA). The level of significance adopted was 5%. Statistical differences were observed in blastocyst production rate: Control: 42.64% and BL-I + ROS: 55.96% (P < 0.05). The remaining embryos (Control n = 163; BL-I + ROS n = 182) were vitrified, warmed, and recultured in SOFaa with 10% FCS for 12 h. After this period, re-expansion rate was determined. We found difference in re-expansion rates of groups (Control: 84.66% and BL-I + ROS: 93.40%; P < 0.05). Additionally, differences were found among total number cell groups: Control: 54.33 and BL-I + ROS: 97.5; P < 0.05). The use of BL-I + ROS is able to better a producing rate of in vitro embryos and improve the cryotolerance. The authors acknowledge FAPESP 2010/18994-4 and 2011/08926-4 for financial support.

scholarly journals In vitro production of bovine embryos

2003 ◽  
Vol 57 (3-4) ◽  
pp. 257-264
Author(s):  
Vojislav Pavlovic ◽  
Jelena Aleksic
Keyword(s):  
Pharmaceutical Products ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Cattle Breeding ◽  
Vitro Fertilization ◽  
Laboratory Conditions ◽  
Female Donor ◽  
Vitro Production ◽  
Bovine Oocytes

Biotechnology is used for the purpose of improving production, and developing animal and pharmaceutical products. In roder to achieve these objectives, it is necessary to manipulate these processes. Reproductive biotechnology can be used independently, or it can be used in connection with other techniques. Thus, for instance, successful culture of embryos in laboratory conditions is a necessary precondition for the production and creation of transgenic and cloned animals. The in vitro process of embryo production is narrowed down to three basic steps: 1. collecting oocytes form a female donor, 2. fertilization of oocytes under laboratory conditions, 3. growth of the embryo in a medium and transfer of the embryo into the recipient. The paper describes the IVP procedure (in vitro production) of bovine embryos; the advantages and shortcomings of this method, as well as possibilities for its application in cattle breeding. This technology is still quite new, so taht both the technique and the mediums are constantly being improved. The technique of fertilizing bovine oocytes, as well as their development in laboratory conditions was discovered in 1980, and the first calf produced using in vitro fertilization (IVF) was born in 1982. IVP implies a series of steps, and if just one of them is not done perfectly, the result is a small number of embryos, or even none at all.

scholarly journals In Vitro Production of Embryos at Research and Development Station for Cattle Breeding Dancu, Iasi – First Bovine Embryos Produced In Vitro in North-Eastern Romania

2017 ◽  
Vol 50 (4) ◽  
pp. 109-117
Author(s):  
S.I. Borş ◽  
Şt. Creangă ◽  
D.L. Dascălu ◽  
Elena Ruginosu ◽  
Mădălina Alexandra Davidescu ◽  
...  
Keyword(s):  
Genetic Gain ◽  
Genetic Improvement ◽  
Selection Intensity ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Cleavage Rate ◽  
Cattle Breeding ◽  
North Eastern ◽  
Vitro Production

AbstractThein vitroproduction (IVP) of bovine embryos increases the selection intensity in cattle and reduces the generation interval, which is very important in the genetic gain. In Romania, this reproductive biotechnology has shown a timid evolution in the last years, although the need for genetic improvement in the area is present. The aim of this paper is to describe the work that resulted in first bovine embryos obtained through IVP in North-Eastern Romania. Oocytes were collected by slashing ovaries from slaughtered cows, matured in a TCM199-based medium and fertilized in TL-based medium microdrops with sperm processed by swim-up procedure. The presumptive embryos were cultured one day in TCM199 and 8 days in SOF-based medium and evaluated in days 7, 8 and 9 after fertilization. We retrieved an average number of 83 usable oocytes/IVF session, which represents 73.8% from the total harvested oocytes. The average number of cleaved embryos was 50.8 per IVF, reflecting an average cleavage rate of 61.2%. An average of 8.6 blastocysts/IVF session was obtained, representing 10.4% of the selected oocytes or 16.9% of the number of cleaved embryos. Although suboptimal, the results were comparable with other reports on IVP in cattle. The adapted IVP protocol, based on maturation with TCM199, fertilization in microdrops of TL and culture of presumptive embryos one day in TCM199 and afterwards in SOF seems to offer acceptable results and will be used for further attempts to produce bovine embryos.

Noninvasive characterization of metabolites secreted in culture media by bovine embryos during in vitro production

Metabolomics ◽  
2016 ◽  
Vol 12 (5) ◽  
Author(s):  
Érika Cristina dos Santos ◽  
Camila Bruna de Lima ◽  
Kelly Annes ◽  
Marcella Pecora Milazzotto
Keyword(s):  
Culture Media ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Noninvasive Characterization ◽  
Vitro Production

Animal factors that influence the in-vitro production of bovine embryos

1999 ◽  
Vol 51 (1) ◽  
pp. 334 ◽  
Author(s):  
H Twagiramungu ◽  
N Morin ◽  
C Brisson ◽  
G Carbonneau ◽  
J Durocher ◽  
...  
Keyword(s):  
Bovine Embryos ◽  
In Vitro Production ◽  
Vitro Production

65 Comparison of SexedULTRA-4M™ X and Y chromosome-bearing semen versus conventional semen for the in vitro production of bovine embryos

2022 ◽  
Vol 34 (2) ◽  
pp. 268
Author(s):  
A. Velázquez-Roque ◽  
H. Álvarez-Gallardo ◽  
M. Kjelland ◽  
M. Pérez-Martínez ◽  
F. Villaseñor-González ◽  
...  
Keyword(s):  
Y Chromosome ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Vitro Production

Evaluation of different culture systems on the in vitro production of bovine embryos

2005 ◽  
Vol 63 (4) ◽  
pp. 1131-1141 ◽  
Author(s):  
Daniela Costa Pereira ◽  
Margot Alves Nunes Dode ◽  
Rodolfo Rumpf
Keyword(s):  
Bovine Embryos ◽  
In Vitro Production ◽  
Culture Systems ◽  
Vitro Production

221 EFFECT OF STAGE OF CORPUS LUTEUM DEVELOPMENT ON THE IN VITRO PRODUCTION OF BOVINE EMBRYOS

2011 ◽  
Vol 23 (1) ◽  
pp. 209
Author(s):  
S. Miyashita ◽  
K. Miyata ◽  
C. Tachibana ◽  
Y. Inaba ◽  
H. Koyama ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Calf Serum ◽  
Cumulus Cells ◽  
Large Mass ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Stage Of Development ◽  
The Mean ◽  
Vitro Production

The objective of this study was to investigate the effect of stage of corpus luteum (CL) development on the in vitro production of bovine embryos. Ovaries were classified according to the expected day of the oestrous cycle based on the morphology of the ovaries. Ovaries with a corpus hemorrhagicum and the remnant of the follicular lumen filled with blood were considered the early luteal stage (Days 2 to 4; Day 0 = day of ovulation, n = 46). Ovaries with a large mass of orange tissue in the CL were classified as the midluteal stage (Days 7 to 10, n = 42). Cumulus–oocyte complexes (COC) were collected by aspiration of 2- to 6-mm follicles. The COC were classified into the following grades: COC with >3 compact layers of cumulus cells and evenly granulated cytoplasm were classified into Grade 1; COC with >3 layers cumulus cells and evenly granulated cytoplasm were classified into Grade 2; COC with partially remaining cumulus cells and abnormal cytoplasm were classified into Grade 3; COC without cumulus cells or those with expanded cumulus cells were classified into Grades 4 and 5, respectively. Grades 1 and 2 COC were in vitro matured for 20 h in TCM-199 supplemented with 5% calf serum and 0.02 mg mL–1 of FSH at 38.5°C under an atmosphere of 5% CO2 in air. Matured COC were inseminated with 5 × 106 sperm for 18 h. Presumptive zygotes were cultured in CR1aa medium supplemented with 5% calf serum at 38.5°C under an atmosphere of 5% O2, 5% CO2, and 90% N2 for 9 days (fertilization = Day 0). The mean number of COC and the proportion of COC classified as Grades 1 and 2 were analysed by ANOVA. Cleavage rates on Day 3 and blastocyst rates on Days 7 to 9 were analysed by a chi-square test. The mean number of recovered oocytes in the early luteal stage (18.7 ± 9.5) was significantly higher (P < 0.05) than the number in the midluteal stage (12.2 ± 5.7). The proportion of Grades 1 and 2 oocytes in the early luteal stage [66.7% (531/789)] was significantly higher (P < 0.01) than that in the midluteal stage [51.6% (252/484)]. The cleavage and blastocyst rates in the early luteal stage [60.9% (181/297) and 32.7% (97/297), respectively] were significantly higher (P < 0.05) than those in the midluteal stage [50.7% (76/150) and 20.7% (31/150) respectively].The present study suggests that the stage of development of the CL in bovine ovaries influences the number of recovered oocytes per ovary and the development of in vitro production of bovine embryos.

The SPOM-adapted IVM system improves in vitro production of bovine embryos

2020 ◽  
Vol 158 ◽  
pp. 277-282
Author(s):  
Gabriela Ramos Leal ◽  
Maria Alice Guimarães Graciosa ◽  
Clara Ana Santos Monteiro ◽  
Renata Pasolini ◽  
Agostinho Jorge dos Reis Camargo ◽  
...  
Keyword(s):  
Bovine Embryos ◽  
In Vitro Production ◽  
Vitro Production
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