In vitro production of sheep blastocysts from IVM-oocytes using frozen semen and oviduct epithelial cell co-culture for IVF

1991 ◽  
Vol 35 (1) ◽  
pp. 214 ◽  
Author(s):  
P. Holm ◽  
B. Irvine ◽  
D.T. Armstrong ◽  
R.F. Seamark
Keyword(s):  
Epithelial Cell ◽  
In Vitro Production ◽  
Frozen Semen ◽  
Oviduct Epithelial Cell ◽  
Vitro Production

Successful use of oviduct epithelial cell coculture for in vitro production of viable red deer (Cervus elaphus) embryos

2005 ◽  
Vol 64 (8) ◽  
pp. 1729-1739 ◽  
Author(s):  
Y. Locatelli ◽  
Y. Cognié ◽  
J.C. Vallet ◽  
G. Baril ◽  
M. Verdier ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Cervus Elaphus ◽  
Red Deer ◽  
In Vitro Production ◽  
Oviduct Epithelial Cell ◽  
Vitro Production

264 EFFECT OF FROZEN SEMEN BATCH ON THE IN VITRO PRODUCTION OF BOVINE EMBRYOS

2010 ◽  
Vol 22 (1) ◽  
pp. 289
Author(s):  
M. B. Fernandes ◽  
T. L. G. Torregrossa ◽  
R. B. Prado ◽  
R. A. Vila ◽  
F. P. Elias ◽  
...  
Keyword(s):  
Embryo Development ◽  
Bos Indicus ◽  
Cumulus Cells ◽  
Development Rate ◽  
In Vitro Production ◽  
Vitro Fertilization ◽  
Significance Level ◽  
Frozen Semen ◽  
Vitro Production

Within an in vitro production controlled system, bulls differ with respect to their semen potential in generating embryos when the variables of maternal effect are minimized (Marquant-le-Guienne and Humblot 1992 Ann. Zootech. 41, 361-370). We have tested the hypothesis that even with this variation among bulls, there is also an intra-bull variation, according to the frozen semen batch used in the in vitro fertilization, identified with the date of ejaculate and its freezing. In an embryo commercial production system, over 12 months, 10 619 viable oocytes were obtained by ultrasound-guided follicular aspiration from 642 Nelore cows (Bos indicus). The oocytes were matured in vitro for 24 h in TCM-199 supplemented with 0.5 μg mL-1 FSH, 50 μg mL-1 LH, and 10% fetal bovine serum. They were then inseminated for 18 hours in IVF-TALP medium, using the semen from 4 bulls (A to D) subdivided into 4 frozen batches (I to IV) and selected by 45/90% Percoll gradient. Putative zygotes surrounded in cumulus cells were transferred in CR2aa medium drops (Rosenkrans and First 1994 J. Anim. Sci. 72, 434-437) for 163 h at 39°C in a humidified atmosphere of 5% CO2 in air. The oocyte distribution, the total number of blastocysts, and the embryo development rate by each bull and respective batch are described in Table 1. The chi-square test was measured with a significance level of P < 0.05 and showed that there is a difference between the used batches of each bull regarding the development rate of blastocysts 163 h after IVF Therefore, there is intra-bull variation in the ability to develop in vitro embryos according to the batch of frozen semen. Table 1.Viable oocytes (VO), total blastocysts (TB), and embryo development rate (%E) by bull and batch used in IVF

scholarly journals Establishing an in vitro production program for buffalo embryos (Bubalus bubalis) in Colombia

2015 ◽  
pp. 4495-4504
Author(s):  
Felipe Gamarra P ◽  
Viviana Rendón V ◽  
Aldemar Chávez R ◽  
Leonardo Perez S ◽  
Walter Cardona-Maya ◽  
...  
Keyword(s):  
Bubalus Bubalis ◽  
Fertilization In Vitro ◽  
Natural Cycle ◽  
Ultrasound Guided ◽  
In Vitro Production ◽  
Production Program ◽  
Frozen Semen ◽  
Sexed Semen ◽  
Vitro Production

Objective. Evaluate the results of the standardization of the in vitro production program of buffalo embryos, using oocytes obtained by ultrasound guided oocyte puncture during the 2012 breeding season in Colombia. Materials and methods. Fifty seven buffalo females were selected for ultrasound guided transvaginal aspiration of follicles, oocytes were identified within follicular fluid, classified and transported to the laboratory and matured in vitro for 18 to 20 hours. Frozen semen of seven Mediterranean bulls were used, motile sperm was obtained using the Percoll technique and oocytes were inseminated with 2 million sperm/ml. Presumptive zygotes were cultured for 6 days, grade 1 embryos obtained were frozen using ethylene glycol. Embryos were transferred to females on day 5 during natural cycle. Results. 97 aspirations were performed on the 57 animals, in 8.2% of the aspirations no oocytes were found. 8 oocytes/aspiration were obtained. Of the 783 oocytes, 92% were classified as viable (721/783) and were fertilized. The cleavage and blastocyst rate were 23% and 19% respectively. 37 embryos were transferred and 11 pregnancies were obtained, confirmed by rectal palpation 60 days after transfer, with a pregnancy rate of 29.7%. Conclusions. The results presented here are comparable with others in literature and shows the feasibility of producing in vitro embryos and pregnancies after the standardization of current protocols, with normal and sexed semen and transfer during natural cycle in buffalo.Key words: Buffaloes, embryo, fertilization in vitro, reproduction (Source: MeSH).

298 THE INFLUENCE OF DISTINCT CENTRAL BULL STATIONS ON THE IN VITRO EMBRYONIC DEVELOPMENT OF NELORE BULLS

2010 ◽  
Vol 22 (1) ◽  
pp. 305
Author(s):  
A. Renzi ◽  
F. P. Elias ◽  
R. A. Vila ◽  
R. B. Lôbo
Keyword(s):  
Calf Serum ◽  
Cumulus Cells ◽  
In Vitro Production ◽  
Freezing And Thawing ◽  
Chi Square ◽  
Frozen Semen ◽  
Chi Square Test ◽  
Vitro Production ◽  
In Vitro Embryonic Development

Reproductive biotechnology is growing worldwide as one of the most important tools of cattle breeding because it accelerates the process of genetic improvement. Most of the embryos produced are obtained using frozen semen from different AI centers. During freezing and thawing of semen, the sperm can be damaged by the rapid and dramatic changes in the physicochemical conditions that occur during cooling and ice formation. It has previously been described that bad management of frozen semen can result in reduced fertilization. This study investigated the influence of different central bull stations on the development of in vitro-produced bovine embryos. We compared semen of 154 Nelore bulls, used for IVF, from 8 different central bull stations (all of which used the same cryopreservation protocol) in the development of blastocysts. The in vitro production of embryos was performed as described: oocytes were collected from the slaughterhouse and matured in TCM-199 + 10% fetal calf serum (FCS) +0.5 μg mL-1 FSH + 50 μg mL-1 LH+ 1 μg mL-1 estradiol, for 24 hat 38.5°C in 5%CO2 in atmospheric air. Viable spermatozoids were obtained by centrifugation in Percoll gradient (45 and 90%), and used for IVF in a concentration of 2 million spermatozoa per mL in TALP + 10 μg mL-1 of heparin medium. After 12 h, the presumptive zygotes were transferred to a CR2+ 10% FCS medium and co-cultured with cumulus cells. After 168 h of IVF, we evaluated the number and stage of cleaved embryos produced with the semen of each bull. Statistical analyses were performed by using the chi-square test. Our results suggest that there are differences among distinct central bull stations in the proportion of embryos that developed into blastocysts and the different stages they hatched. FAPESP, CNPq, PROEX, FAEPA.

230 OVUM PICKUP - IN VITRO PRODUCTION EFFICIENCY FOR CONSERVATION OF THE FLAMENGA CATTLE BREED

2011 ◽  
Vol 23 (1) ◽  
pp. 213
Author(s):  
F. C. Zago ◽  
S. Gaudêncio Neto ◽  
L. H. Aguiar ◽  
L. F. Schutz ◽  
F. Forell ◽  
...  
Keyword(s):  
Yield Potential ◽  
Holstein Cows ◽  
Recovery Efficiency ◽  
Southern Brazil ◽  
In Vitro Production ◽  
Developmental Potential ◽  
Embryo Yield ◽  
Frozen Semen ◽  
Vitro Production

The introduction of more specialised and productive dairy breeds into commercial herds in recent decades has caused a gradual loss of interest in dual-purpose cattle breeds, such as the Flamenga. Because of its importance to biodiversity, the high risk of loss of this small Brazilian herd, located in one geographic region in Southern Brazil, justifies the need for and efforts in genetic conservation of the Flamenga breed. The ovum pickup (OPU) procedure is a potential tool for such conservation purposes. Thus, the aim of this study was to evaluate the oocyte and embryo yield potential of the Flamenga breed, as compared with Holstein control counterparts, after in vivo oocyte retrieval followed by in vitro embryo production (IVP). Four Flamenga and 4 Holstein multiparous milking cows from the same herd were subjected to weekly ultrasound-guided OPU sections for 10 consecutive weeks. The number of visible follicles per ovary per female was recorded before the aspiration of ≥3-mm follicles. Retrieved cumulus–oocyte complexes (COC) were morphologically graded according to the method of (Leibfried and First 1979 J. Anim. Sci. 48, 76–86), and viable COC (Grades I, II, and III) from both breeds and from slaughterhouse ovaries (IVP controls) were used for the IVP of embryos, based on our established procedures (Vieira et al. 2002 Cryobiology 45, 91–94). Frozen semen from the same Flamenga breed bull was used in all groups. Quantitative (visible and aspirated follicles, collected COC), and qualitative (COC recovery efficiency, cleavage and blastocyst rates) data were compared between breeds and IVP controls (when pertinent) by Student’s t-test and chi-square test, respectively, for P < 0.05. Results are presented in Table 1. The pools of visible and aspirated follicles were similar between breeds. However, the COC recovery efficiency was higher for Flamenga than for Holstein females. In vitro embryo development was also different between breeds, with cleavage rate being significantly lower for Holsteins than for Flamenga and IVP controls. Blastocyst rates also differed between treatments. Nonetheless, development to the blastocyst stage, when based on cleavage, was similar between Flamenga and Holstein embryos, but with both breeds being lower than the IVP controls. Oocyte quality was likely the cause of such differences in embryo developmental potential between groups, with Flamenga females yielding more oocytes and IVP embryos than Holstein cows. In summary, the use of OPU-IVP procedures was demonstrated to be suitable for the preservation of the Flamenga breed in Southern Brazil. Table 1.Follicular pool and oocyte and embryo yield between Flamenga and Holstein cows following ovum pickup–in vitro production (IVP) procedures This study was supported by CAPES and CNPq/Brazil.

scholarly journals 144 EFFECT OF ANTRAL FOLLICLE COUNT IN BEEF HEIFERS ON IN VITRO FERTILIZATION/PRODUCTION

2017 ◽  
Vol 29 (1) ◽  
pp. 180
Author(s):  
C. C. Chase ◽  
R. A. Cushman ◽  
A. K. McNeel ◽  
E. C. Wright-Johnson ◽  
G. A. Perry ◽  
...  
Keyword(s):  
Antral Follicle ◽  
Antral Follicle Count ◽  
Blastocyst Stage ◽  
In Vitro Production ◽  
Blastocyst Development ◽  
University Of Florida ◽  
Vitro Fertilization ◽  
Frozen Semen ◽  
Vitro Production

Our objective has been to compare the IVF and in vitro production (IVP) of embryos from low and high antral follicle count (AFC) heifers. This is the fourth year of the study with years 1 to 3 reported individually. For this report, we add data for the fourth year and present a combined analysis (years 1 to 4) for the first time. Each year, AFC was determined on ~120 Angus heifers using transrectal ultrasonography. Ten heifers with the lowest AFC and 10 heifers with the highest AFC and all with evidence of oestrous cyclicity were synchronized with two 5-mL injections of PGF2α 11 days apart. Half were harvested on Day 5 to 6 and half on Day 15 to 16 of the oestrous cycle. The IVF procedure was slightly modified each year. For year 4, the IVF procedure included protocols for semi-defined media and was as described (IVP Protocol, P. J. Hansen’s Laboratory, University of Florida). Cumulus-oocyte complexes (COC) from follicles less than 8 mm in diameter were cultured in maturation medium (5% CO2; 38.5°C) for 24 h. Matured COC were fertilized using thawed frozen semen from a bull that was purified using isolate. Motile spermatozoa were added to COC in fertilization medium at a final concentration of 1 × 106 spermatozoa per mL. About 24 h later, presumptive zygotes were placed in micro drops of development medium under oil, and cultured (5% CO2; 5% O2; balance N2; 38.5°C). On Day 3 and 8 after fertilization, cleavage and blastocyst development rates, respectively, were assessed. Data were analysed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC, USA) and the model included the effects of year (1 to 4), group (high or low AFC), and their interaction. The year × group interaction was not significant (P > 0.10). Low AFC heifers, compared with high AFC heifers, had fewer numbers of COC (P < 0.0001; 12.8 ± 1.83 v. 31.9 ± 1.86), fewer numbers of COC that cleaved (P < 0.0001; 8.0 ± 1.38 v. 21.6 ± 1.40), and fewer numbers of COC that developed to the blastocyst stage (P < 0.0001; 1.7 ± 0.58 v. 5.7 ± 0.58). Year affected the numbers of COC that cleaved (P < 0.003) and the numbers of COC that developed to the blastocyst stage (P < 0.0001). Year also influenced the percentage of COC that cleaved (P < 0.0002) and the percentage of COC that developed to blastocysts (P < 0.0001). Group (AFC) did not influence (P > 0.19) the percentage of COC that cleaved (61.2 ± 2.83 v. 66.4 ± 2.83%, for low v. high AFC, respectively). Low AFC heifers had a lower (P < 0.002) percentage of COC that developed to blastocysts (10.3 ± 1.52%) than high AFC heifers (17.6 ± 1.52%). These results indicate that high AFC heifers, compared to low AFC heifers, have more COC recovered, more COC cleaved, and more COC developed to the blastocyst stage. The percentage of COC cleaved did not differ between AFC groups; however, the percentage of COC that developed to the blastocyst stage was greater for high than low AFC heifers. This suggests a potential advantage in maternal to embryonic transition for high compared with low AFC heifers.

THE IN VITRO PRODUCTION OF ANDROGENS BY FOLLICULAR TISSUE OF RABBITS

1964 ◽  
Vol 47 (2) ◽  
pp. 306-313 ◽  
Author(s):  
Denis Gospodarowicz
Keyword(s):  
In Vitro Production ◽  
Vitro Production

ABSTRACT Incubation in vitro of rabbit follicles in separate experiments with dehydroepiandrosterone-14C (DHEA-14C), progesterone-14C and pregnenolone-3H in the presence of FSH gave the following results: 39 % of the radioactivity of DHEA-14C is converted to androstenedione and testosterone, while only 3 % of the radioactivity of either progesterone-14C or pregnenolone-3H is found in the androgen fraction. From the ratio of testosterone to androstenedione formed from the three precursors, the results are interpreted to mean that DHEA and pregnenolone, and not progesterone, are precursors of androgens in the follicle.

Oxygenation of 18-hydroxycorticosterone as the final reaction for aldosterone biosynthesis

1984 ◽  
Vol 107 (3) ◽  
pp. 395-400 ◽  
Author(s):  
Itaru Kojima ◽  
Etsuro Ogata ◽  
Hiroshi Inano ◽  
Bun-ichi Tamaoki
Keyword(s):  
Carbon Monoxide ◽  
Hydroxysteroid Dehydrogenase ◽  
Dependent Manner ◽  
In Vitro Production ◽  
Mitochondrial Preparation ◽  
Final Reaction ◽  
Vitro Production ◽  
Dose Dependent ◽  
Cytochrome P 450

Abstract. Incubation of 18-hydroxycorticosterone with the sonicated mitochondrial preparation of bovine adrenal glomerulosa tissue leads to the production of aldosterone, as measured by radioimmunoassay. The in vitro production of aldosterone from 18-hydroxycorticosterone requires both molecular oxygen and NADPH, and is inhibited by carbon monoxide. Cytochrome P-450 inhibitors such as metyrapone, SU 8000. SU 10603, SKF 525A, amphenone B and spironolactone decrease the biosynthesis of aldosterone from 18-hydroxycorticosterone. These results support the conclusion that the final reaction in aldosterone synthesis from 18-hydroxycorticosterone is catalyzed by an oxygenase, but not by 18-hydroxysteroid dehydrogenase. By the same preparation, the production of [3H]aldosterone but not [3H]18-hydroxycorticosterone from [1,2-3H ]corticosterone is decreased in a dose-dependent manner by addition of non-radioactive 18-hydroxycorticosterone.

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