Effect of time during transport of excised mare ovaries on oocyte recovery rate and quality after in vitro maturation

1999 ◽  
Vol 52 (5) ◽  
pp. 757-766 ◽  
Author(s):  
F. Guignot ◽  
J. Bezard ◽  
E. Palmer
Keyword(s):  
Recovery Rate ◽  
In Vitro Maturation ◽  
Oocyte Recovery

243 EFFECT OF OOCYTE RECOVERY TECHNIQUES ON IN VITRO MATURATION OF SWINE OOCYTES

2009 ◽  
Vol 21 (1) ◽  
pp. 219
Author(s):  
F. R. O. de Barros ◽  
M. G. Marques ◽  
M. D. Goissis ◽  
M. A. Peres ◽  
M. P. Milazzotto ◽  
...  
Keyword(s):  
Follicular Fluid ◽  
Recovery Rate ◽  
In Vitro Maturation ◽  
Metabolic Stress ◽  
Water Bath ◽  
Epifluorescence Microscopy ◽  
Nuclear Maturation ◽  
Oocyte Recovery ◽  
Centrifuge Tube

The aim of this study was to compare 2 different techniques to obtain swine oocytes from abattoir ovaries. Ovaries were washed in saline at 35°C and submitted to slashing or aspiration, simultaneously. For the slashing group, ovaries were held with a hemostat inside a beaker containing 35 mL of HEPES-buffered Tyrode’s media (HbT) and follicles (2–6 mm) were incised with a scalpel. For every 5 slashed ovaries, HbT-containing follicular fluid was transferred to 50-mL centrifuge tubes. For the aspiration group, follicles (2–6 mm) were aspirated using an 18-gauge needle and a 5-mL syringe. The follicular fluid of each ovary was transferred to a 50-mL centrifuge tube. Tubes from both techniques were placed in a water bath at 35°C for 15 min to allow settling of the cumulus–oocyte complexes (COC). The supernatant was removed and the sediment was resuspended in HbT and placed in water bath at 35°C for an additional 15 min. The sediment was resuspended in 15 mL of HbT and COC were recovered under stereomicroscopy. Oocytes were in vitro matured for 44 h in TCM-199 added with 10% porcine follicular fluid (PFF) and hormones (LH and FSH) at 38.5°C, 5% CO2 and high humidity. The oocyte recovery rate of each technique was determined by the ratio between the number of COC and ovaries used. To verify nuclear maturation by epifluorescence microscopy (Zeiss), oocytes were fixed, permeabilized, and incubated in 10 μg mL–1 of RNAse for 30 min and in 10 μg mL–1 of propidium iodide for 10 min. Heat shock protein 70 (HSP70) content was assessed as described in Kawarsky and King (2001 Zygote 9(3), 39–50) to verify the metabolic stress. Data were analyzed by ANOVA and Tukey’s test using the software Statistica for Windows. A level of 5% was considered significant in all assessments. The oocyte recovery rate (COC/ovary) was higher for the slashing group (2.665 ± 0.38) compared with the aspiration group (1.762 ± 0.15). The percentage of oocytes that reached the germinative vesicle (GV) stage (h 0 of maturation) did not differ between groups (100 ± 0 and 86.66 ± 13.36, slashing and aspiration group, respectively). The same was observed for the percentage of oocytes that reached the metaphase II stage (MII, after 44 of maturation; 79.99 ± 9.74 and 96.00 ± 4.00, slashing and aspiration group, respectively). Moreover, no difference at pixel quantification of HSP70 was observed between groups (256.50 ± 42.42 and 238.61 ± 71.18, slashing and aspiration group, respectively). In conclusion, the slashing procedure provided a better oocyte recovery rate compared with the aspiration of ovaries. This technique does not affect nuclear maturation, because no differences were observed regarding the percentage of oocytes that reached the GV and MII stages. In addition, it does not affect HSP70 content, suggesting that the slashing of ovaries does not increase the basal stress of oocytes in an in vitro-maturation system.

Effect of administering a crude equine gonadotrophin preparation to mares on follicular development, oocyte recovery rate and oocyte maturation in vivo

Reproduction ◽  
2000 ◽  
pp. 351-360 ◽  
Author(s):  
I Bruck ◽  
J Bezard ◽  
M Baltsen ◽  
B Synnestvedt ◽  
I Couty ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Recovery Rate ◽  
In Vitro Maturation ◽  
Follicular Development ◽  
Vitro Fertilization ◽  
Nuclear Maturation ◽  
Oocyte Yield ◽  
Oocyte Recovery

In mares, the shortage of oocytes and the variability in nuclear maturation at a certain time of the oestrous cycle hinders the optimization of methods for in vitro maturation and in vitro fertilization. Increasing the number of small-to-medium-sized follicles available for aspiration in vivo may increase the overall oocyte yield. The aims of the present study were to investigate whether administration of crude equine gonadotrophins affects follicular development, oocyte recovery rate, in vivo oocyte maturation and follicular concentrations of meiosis-activating sterols. During oestrus, all follicles >/= 4 mm were aspirated from 19 pony mares (first aspiration: A1). Over the next 8 days, the mares were treated daily with either 25 mg crude equine gonadotrophins (n = 10) or physiological saline (n = 9). Between day 1 and day 8, follicular growth was monitored by ultrasonography. On day 8, all follicles >/= 4 mm were evacuated (second aspiration: A2) and nuclear maturation of the recovered oocytes was assessed after orcein staining. Follicular growth between A1 and A2, as well as the number and size of follicles at A2 were similar for control mares and mares treated with crude equine gonadotrophins. The oocyte recovery rates at A1 and A2 were similar. At A2, the oocyte recovery rate and oocyte maturation in vivo were not affected by treatment with crude equine gonadotrophins. The number of expanded cumulus oophorus complexes recovered from follicles </= 29 mm was significantly higher at A1 than at A2. The number of oocytes at the germinal vesicle stage was significantly higher at A2 (41.5%) than at A1 (17.8%). Meiosis-activating sterols (FF-MAS and T-MAS) were identified in follicular fluid recovered at A2. Follicular concentrations of FF-MAS and T-MAS were unaffected by treatment with crude equine gonadotrophins. The present study demonstrates that follicular aspiration during oestrus allowed a new follicular population to develop and resulted in a higher degree of synchronization of oocyte development with respect to cumulus expansion and nuclear maturation. The availability of a more homogeneous population of oocytes might facilitate a better optimization of in vitro maturation and in vitro fertilization techniques in mares. Administration of crude equine gonadotrophins during early dioestrus did not affect the growth of small follicles, the oocyte yield after aspiration or oocyte maturation in vivo.

In vitro embryo production using ovaries removed from culled cows

2001 ◽  
Vol 81 (4) ◽  
pp. 589-591 ◽  
Author(s):  
G. Giritharan ◽  
R. Rajamahendran
Keyword(s):  
Dairy Cows ◽  
Key Words ◽  
Estrous Cycle ◽  
Recovery Rate ◽  
In Vitro Maturation ◽  
Follicle Stimulating Hormone ◽  
Embryo Production ◽  
Oocyte Recovery ◽  
In Vitro Embryo Production

The objective was to utilize in vitro maturation and fertilization to maximize embryo production from culled dairy cows. Ovaries were removed (via a colopotomy technique) from culled Holstein cows (n = 21) 2 d after standing estrus (T1), 2 d after treatment with 40 mg follicle stimulating hormone (FSH) given at standing estrus (T2), or at an unknown stage of the estrous cycle (T3).Treatment with FSH increased (P < 0.05) oocyte recovery rate (mean ± SEM, 14.6 ± 2.2, 25.4 ± 3.4, and 11.1 ± 2.6 oocytes ovary–1), for groups T1, T2 and T3, respectively, and the number of blastocysts obtained (1.6 ± 0.5, 4.4 ± 0.4 and 1.6 ± 0.6 blastocysts ovary–1). Culled dairy cows of high genetic merit are potential embryo donors, with an average of six to eight transferable embryos obtained from each FSH-treated cow. Key words: FSH treatment, oocyte recovery, IVM, IVF, culled cows

scholarly journals L-OPU in Goat and Sheep—Different Variants of the Oocyte Recovery Method

Animals ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 658
Author(s):  
Jarosław Wieczorek ◽  
Jurij Koseniuk ◽  
Maria Skrzyszowska ◽  
Mirosław Cegła
Keyword(s):  
In Vitro Maturation ◽  
Hormonal Stimulation ◽  
Recovery Method ◽  
Vitro Fertilization ◽  
Minimal Invasiveness ◽  
Animal Reproduction ◽  
Oocyte Recovery ◽  
Maturation In Vitro ◽  
Invasive Techniques

The laparoscopic method of recovering oocytes in goats and sheep is one of the minimally invasive methods used in the biotechnology of animal reproduction. It allows for good quality oocytes that are suitable for in vitro maturation and fertilization to be recovered. The limitation of using the laparoscopic ovum pick-up (L-OPU) method in goat and sheep is its changing effectiveness and the lack of repeatability of results, as well as the varying effectiveness of different variants of the method. Therefore, it is necessary to develop effective non-invasive techniques allowing for multiple good quality oocyte recovery that would be suitable for in vitro maturation and fertilization. In this study, four different L-OPU variants were described in goats and sheep. Various techniques of recovering oocytes were discussed, including the techniques of conducting the operation, various tools for recovering oocytes, and different plans of hormonal stimulation. Recovery rates were 35% (Variant I), 57% (Variant II), 72% (Variant III), and 67% (Variant IV). After evaluation, 94% (both Variant I and II), 93% (Variant III), and 84% (Variant IV) of the oocytes were qualified for in vitro maturation. The results of the study show that the proposed technique of laparoscopic recovery of oocytes allows a sufficient number of ovarian cells suitable for in vitro culture to be obtained and as a consequence it makes them useful in in vitro maturation/in vitro fertilization (IVM/IVF) programs or cloning. The method allows for a fast and effective conduct of the operation in a living donor with minimal invasiveness while preserving the excellent condition of animals.

scholarly journals In vivo ultrasound guided transvaginal oocyte collection in the cow

2018 ◽  
Vol 49 (3) ◽  
pp. 195
Author(s):  
G. S. AMIRIDIS (Γ.Σ. ΑΜΟΙΡΙΔΗΣ) ◽  
M. SALAHEDDINE ◽  
I. A. JEFFCOATE ◽  
E. VAINAS (Ε. ΒΑΪΝΑΣ) ◽  
L. ROBERTSON
Keyword(s):  
Recovery Rate ◽  
Transvaginal Ultrasound ◽  
Oestrous Cycle ◽  
Ultrasound Guided ◽  
Follicular Aspiration ◽  
Oocyte Recovery ◽  
Oocyte Collection ◽  
Significant Difference

This paper describes the results of the in vivo ultrasound guided follicular aspiration for ovum pick υρ (OPU) in the cow. Twelve non pregnant dry cows aged 4-6 years were used in this experiment. Eight cows underwent OPU during three successive oestrous cycles and another four cows were used as controls having only transvaginal ultrasound scanning of their ovaries. Oocyte collection took place three times during the luteal phase of each natural oestrous cycle (days 3-4,9-11 and 14-17). The content of 326 follicles with a diameter of 4-15mm was aspirated and 104 oocytes were collected (recovery rate 31.9% or 1.55 oocytes per cow and session). The oocyte recovery rate increased after the first three sessions (from 13.04% to 35.0%) and reached levels of υρ to 52.6%. More follicles were aspirated on days 9-11 (133 follicles 40.8%) compared to 111 (34%) follicles on days 14-17 and 82 (25%) on days 3-4) (P<0.05). The evaluation of the collected oocytes revealed that 60 oocytes (57.7%) were suitable for further in vitro manipulation. Neither the origin of the oocyte (left or right ovary) nor the stage of the oestrous cycle affected the recovery rate or the quality of the collected oocytes. There was no significant difference either in the length of the oestrous cycle between the experimental animals and the controls (21.6± 1.4 vs. 22.37±1.0 respectively), or in plasma progesterone concentration in daily collected blood samples from the animals of the two groups. The results of this study are compared to those from the international literature and to the results from endoscopical methods for oocyte recovery. The feasibility of application of this technique to projects designed to improve the genetic merit of cows is discussed.

scholarly journals Comparative seasonal assessment on the quality of Black Bengal goat oocytes in view of in vitro embryo production

2020 ◽  
Vol 5 (3) ◽  
pp. 78-87
Author(s):  
Shikha Sarkar ◽  
Md Saiful Islam ◽  
Md Enayet Kabir ◽  
Falguni Dadok ◽  
Md Abdullah Al Zaber ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Recovery Rate ◽  
In Vitro Maturation ◽  
Winter Season ◽  
Excellent Opportunity ◽  
Different Seasons ◽  
In Vitro Embryo Production ◽  
Black Bengal Goat

In vitro maturation (IVM) of oocytes provide an excellent opportunity for cheap and abundant embryos for carrying out animal improvement. With the aim for studying the quality of oocyte in different seasons of Black Bengal goat, both right and left ovaries were collected from the slaughter houses. For each of the specimens, gross parameters such as right, left, corpus luteum (CL)-present and corpus luteum (CL)-absent group were evaluated on the basis of weight (g), length (cm), width (cm), follicles aspirated and number and state of cumulus-oocyte-complexes (COCs), normal COCs and abnormal COCs, the effect of season on cumulus oocyte complexes (COCs) of goat oocytes. Our study revealed that the average number of follicles and oocytes recovery rate were higher in summer than in winter season and recovery rate of grade A and B oocytes were higher in summer than that of in winter and recovery rate of grade C and D were higher in winter than that of in summer. In the present study a significantly greater number of oocytes per ovary were recovered from ovaries without a corpus luteum than from ovaries with a corpus luteum. So the summer was the best season for recovery of COCs which have a worthy competence to be matured in vitro. Asian Australas. J. Biosci. Biotechnol. 2020, 5 (3), 78-87

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