Improvement of cortical granules migration and in vitro embryo production of vitrified bovine oocyte by 9-cis retinoic acid

In this study the effect of increasing ethylene glycol (EG) concentrations in the vitrification media (VM) and its relation with oocyte activation and in vitro embryo production were evaluated. Cumulus-oocytes complexes (COC) were matured in vitro for 22 h and then partially denuded by gently pipetting with hyaluronidase, and randomly assigned to 4 treatments: T1 = control group; T2 = COC exposed to 10% EG + 10% DMSO (VM1), 20% EG + 20% DMSO (VM2); T3 = 15% EG + 5% DMSO (VM1), 30% EG + 10% DMSO (VM2); T4 = 20% EG + 0% DMSO (VM1), 40% EG + 0% DMSO (VM2). Exposition to VM1 and VM2 lasted 3 min and 30 s, respectively. After treatment COC were incubated in maturation medium up to 24 h. In Experiment 1 COC were cultured for 24 h in fertilization TALP supplemented with 50 μg mL-1 of heparin, then completely denuded and cultured 24 h in CR1-aa. After this, oocytes were stained with bisbenzimide (Hoechst 33342) and the number of nuclei and cells were recorded. Structures presenting 2 cells or 2 nuclei were considered as activated oocytes. In Experiment 2 matured COC exposed to cryoprotectants as in Experiment 1 were fertilized and cultured for 6 days as previously described (Mucci et al. 2006). Variables analyzed included oocyte activation, cleavage rate at 48 h, and percentage of viable embryos at Day 7. Data were analyzed by PROC GENMOD (SAS Institute Inc., Cary, NC, USA). Activated oocytes percentage did not differ between EG concentrations in VM and were higher (T2, 24.7%, n = 101; T3, 25.0%, n = 96; T4, 30.2%, n = 119) compared with controls (T1, 9.8%, n = 61). In Experiment 2 no differences were found in cleavage rates (T1, 81.9%, n = 68; T2, 87%, n = 67; T3, 85.9%, n = 61; T4, 84.2%, n = 64) and Day 7 percentage of viable embryos (T1, 34.9%, n = 29; T2, 28.6%, n = 22; T3, 26.8%, n = 19; T4, 27.6%, n = 21) in treated COC. The exposition of COC to cryoprotectants per se could trigger oocyte activation in the range of 10 to 40%. Thanks toAdriana Cano (Instituto Nacional de Tecnología Agropecuaria) for contributions in statistical analysis.

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The effects of ceftiofur sodium (Naxcel) on bovine oocyte and preimplantation embryonic development assessed by in vitro embryo production techniques

Journal of Veterinary Pharmacology and Therapeutics ◽

10.1046/j.1365-2885.1998.00116.x ◽

1998 ◽

Vol 21 (2) ◽

pp. 92-98 ◽

Cited By ~ 2

Author(s):

G.R. Holyoak ◽

S. Wang ◽

G. Liu ◽

T.J. Bunch ◽

R.C. Evans ◽

...

Keyword(s):

Embryonic Development ◽

Bovine Oocyte ◽

Embryo Production ◽

Production Techniques ◽

In Vitro Embryo Production ◽

Ceftiofur Sodium

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Bovine Oocyte Meiotic Inhibition Before In Vitro Maturation and Its Value to In Vitro Embryo Production: Does it Improve Developmental Competence?

Reproduction in Domestic Animals ◽

10.1111/j.1439-0531.2011.01924.x ◽

2011 ◽

Vol 47 (4) ◽

pp. 687-693 ◽

Cited By ~ 23

Author(s):

S Bilodeau-Goeseels

Keyword(s):

In Vitro Maturation ◽

Developmental Competence ◽

Bovine Oocyte ◽

Embryo Production ◽

In Vitro Embryo Production

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Laparoscopic Ovum Pick-Up Followed by In Vitro Embryo Production and Transfer in Assisted Breeding Programs for Ruminants

Animals ◽

10.3390/ani11010216 ◽

2021 ◽

Vol 11 (1) ◽

pp. 216

Author(s):

Hernan Baldassarre

Keyword(s):

Pregnancy Rate ◽

Commercial Application ◽

Embryo Production ◽

Breeding Programs ◽

Sheep And Goats ◽

Embryo Recovery ◽

Marker Selection ◽

Holstein Calves ◽

In Vitro Embryo Production

The potential of laparoscopic ovum pick-up (LOPU) followed by in vitro embryo production (IVEP) as a tool for accelerated genetic programs in ruminants is reviewed in this article. In sheep and goats, the LOPU-IVEP platform offers the possibility of producing more offspring from elite females, as the procedure is minimally invasive and can be repeated more times and more frequently in the same animals compared with conventional surgical embryo recovery. On average, ~10 and ~14 viable oocytes are recovered by LOPU from sheep and goats, respectively, which results in 3–5 transferable embryos and >50% pregnancy rate after transfer. LOPU-IVEP has also been applied to prepubertal ruminants of 2–6 months of age, including bovine and buffalo calves. In dairy cattle, the technology has gained momentum in the past few years stemming from the development of genetic marker selection that has allowed predicting the production phenotype of dairy females from shortly after birth. In Holstein calves, we obtained an average of ~22 viable oocytes and ~20% transferable blastocyst rate, followed by >50% pregnancy rate after transfer, declaring the platform ready for commercial application. The present and future of this technology are discussed with a focus on improvements and research needed.

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