In Vitro Fertilization of Bovine Oocytes Matured In Vitro

1997 ◽  
Vol 1987 ◽  
pp. 28-28
Author(s):  
K.H. Lu ◽  
I. Gordon ◽  
M.P. Boland ◽  
T.F. Crosby
Keyword(s):  
Embryo Transfer ◽  
Normal Development ◽  
Present Report ◽  
Blastocyst Stage ◽  
Laboratory Procedure ◽  
Commercial Exploitation ◽  
Vitro Fertilization ◽  
Stage Of Development ◽  
Bovine Oocytes

The development of an efficient laboratory procedure which would enable cattle ovarian oocytes to be matured in vitro, fertilized and cultured in vitro to the blastocyst stage of development could have important practical and scientific implications. The commercial exploitation of certain embryo transfer techniques applicable in cattle (eg., twinning by embryo transfer) might be facilitated by the development of such a procedure and there would be many advantages to having a cheap source of embryos available for research purposes. The present report deals with some of the studies recently carried out in this laboratory aimed at utilising follicular oocytes recovered from the ovaries of cattle slaughtered for beef at the abattoir. Such studies have been undertaken over a period of almost twenty years, starting with the work of Sreenan (1968)* but it now realised that the oocytes of farm mammals are incapable of normal development until after the completion of complex changes during maturation.

154 COMPARISON OF DIFFERENT CULTURE MEDIA AND INCUBATION METHODS ON CULTURING MURINE EMBRYOS IN VITRO USING STRAW AS A RECEPTACLE

2017 ◽  
Vol 29 (1) ◽  
pp. 185
Author(s):  
L. R. Madzhie ◽  
M. A. Raseona ◽  
L. P. Nethenzheni ◽  
O. Ajao ◽  
M. L. Mphaphathi ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Culture Media ◽  
Uv Light ◽  
Fertilization Rate ◽  
Blastocyst Stage ◽  
Blastocyst Formation ◽  
Vitro Fertilization ◽  
Stage Of Development ◽  
Murine Embryos

In vitro fertilization in the straw system might increase the efficiency of fertilization and the quality of blastocyst formation as compared with micro-drops-IVF systems. The aim of the study was to in vitro fertilize mouse oocytes and culture the resulting zygotes in bi-gas incubator and in a goat vagina and compare the in vitro embryo developmental stages in TCM-199 and Ham’s F10 culture media until the blastocyst-stage of development. F1 generations (Balb C × C57) were used to harvest oocytes and spermatozoa. The fresh sperm were capacitated in different incubation methods (bi-gas incubator and in the vagina of a goat). A volume of 2–4 µL of Ham’s F10 containing capacitated sperm (~8 × 106 per mL) were placed into Ham’s F10 fertilization drops under the oil, containing 10 oocytes and penicillamine, hypotaurine, and epinephrine for enhancing sperm motility and penetration of oocytes. The same procedure was used with the TCM-199 medium and IVF drops without oil (both TCM-199 and Ham’s F10) for straw filling. The presumptive embryos in Ham’s F10 and TCM-199 were divided into different groups: first group were cultured in micro-drops, second group the embryos were aspirated in semen straws and placed in the incubator (incubator straws) for culture, and other straws were covered with a sponge and inserted in the vagina of a goat (vaginal straws) for culture. The resulted blastocysts were stained using Hoechst 33528 solution and blastomeres were counted on a fluorescent UV light inverted microscope at 400× magnification (Nikon Eclipse TI, Narishige Co., Ltd., Amityville, NY, USA). The results were analysed by 2 × 2 factorial designs and Student’s t-test was used to separate the mean. There was no statistical difference (P > 0.05) between the media and incubators on the stage of murine embryo development. The overall fertilization rate was 94 to 99%. The incubator straws with Ham’s F10 (80.5%) had the highest rate of embryos that reached the blastocyst stage, followed by incubator straws with TCM-199 (77.0%), and vaginal straws with Ham’s F10 (60.0%) had the lowest rate of embryos that reached the blastocyst stage. The overall mean number of blastomeres in the blastocyst stage of the embryos ranged from 85 ± 9 to 90 ± 9 cells in all receptacles and incubators. It was concluded that the fertilization and culturing of murine embryos are possible in straws incubated in a bi-gas incubator and in the goat vagina as an alternative method of fertilizing oocytes and culturing murine embryos. In addition, Ham’s F10 and TCM-199 can both be used to fertilize oocytes and culture murine embryos until blastocyst formation embryo in vitro, incubated in a bi-gas incubator or in the vagina.

scholarly journals Uterine Factors Modify the Association Between Embryo Transfer Depth and Clinical Pregnancy: A Retrospective Study in 7849 Fresh Transfer In Vitro Fertilization Cycles

2021 ◽  
Author(s):  
Xiaohua Sun ◽  
Jiali Cai ◽  
Lanlan Liu ◽  
Haixiao Chen ◽  
Xiaoming Jiang ◽  
...  
Keyword(s):  
Retrospective Study ◽  
Embryo Transfer ◽  
Endometrial Thickness ◽  
Patient Characteristics ◽  
Blastocyst Stage ◽  
Clinical Pregnancy ◽  
Uterine Contractility ◽  
Vitro Fertilization ◽  
Fresh Transfer

Abstract The embryo position is supposed to affect implantation following embryo transfer. However, embryo dislodging caused by uterine contraction may occurred after transfer. The retrospective study was to investigated whether the factors associated with uterine contractility, such as endometrial thickness and progesterone elevation, affect the association between embryo position and implantation. A total of 7849 fresh transfer cycles on conventional stimulation in a single IVF centre during the period 2013–2015 was reviewed. Patients were categorized according to quartiles of embryo-fundus distance (≤9, 9.1-11, 11.1-14, ≥1.4 mm, respectively). Adjusted for confounding factors, the odds ratio (OR) (95%CI) for clinical pregnancy was 0.90 (0.79-1.02), 0.86 (0.74-0.99) and 0.70 (0.60-0.82) respectively in quartiles 2 through 4, comparing with quartile 1. However, ORs were significantly increased when endometrial thickness was < 8 mm. The ORs comparing quartiles 2 through 4 with quartile 1 increased 1.96 (95%: 1.33-2.90), 1.20 (95%: 0.78-1.87) and 1.98 (95%: 1.20-3.26) fold respectively in cycles with an endometrial thickness < 8 mm than in cycles with a normal endometrial thickness (8-11 mm). Elevated progesterone on the day of hCG and blastocyst stage transfer reduced the ORs. Our data suggested an interaction between patient characteristics and embryo transfer techniques.

185 NONSURGICAL EMBRYO TRANSFER FOR PRODUCTION OF PIGS BY IN VITRO FERTILIZATION AND SOMATIC CELL NUCLEAR TRANSFER

2010 ◽  
Vol 22 (1) ◽  
pp. 251
Author(s):  
J.-G. Yoo ◽  
M.-R. Park ◽  
H.-N. Kim ◽  
Y.-G. Ko ◽  
J.-Y. Lee ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Embryo Transfer ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Blastocyst Stage ◽  
Fibroblast Cells ◽  
Vitro Fertilization ◽  
Miniature Pigs ◽  
Donor Cells

Instead of surgical embryo transfer (ET) in the pig, nonsurgical ET is a hopeful method to increase the efficiency of biotechnology applications such as cloning and transgenesis. In this study, we conducted surgical and nonsurgical ET methods after somatic cell nuclear transfer (SCNT) with MHC miniature pig cells to find out the best condition for production of cloned miniature pigs. Ovaries were obtained from prepubertal crossbred gilts at a local slaughterhouse. Oocytes were matured for 40 to 44 h at 38.5°C under 5% CO2 in air. As donor cells, fibroblast cells were cultured from ear skin tissue of 8-month-old MHC inbred miniature pigs. Fibroblast cells were cultured, passaged (3 to 8 passages), and used as donor cells for NT. After the enucleation and injection process, eggs were held in TCM-199. For fusion, 2 DC pulses of 1.2 kV cm-1 were applied for 30 μs. Both IVF and SCNT embryos were cultured in PZM-3 medium. After IVF, 84.9% (411/484) of embryos cleaved and 27.3% (132/484) of embryos reached the blastocyst stage. In the SCNT group, 80.8% (231/286) of eggs fused and 25.9% (60/286) of embryos developed to blastocysts. For surgical ET, approximately 200 SCNT embryos were transferred into oviducts of each synchronized recipient. For nonsurgical ET, embryos were cultured in PZM-3 for 6 days after SCNT and IVF, and then good quality blastocyst stage embryos were selected for ET. The pregnancy status of recipients at Day 30 was determined by ultrasound scanning. Using Day 30 of gestation as an endpoint, the nonsurgical ET method (47.3%, 9/19) had a similar pregnancy rate as the surgical ET method (56.5%, 13/23). Further study is needed to optimize the nonsurgical ET method especially for SCNT eggs. This work received grant support from the Agenda Program (no. 200901FHT010305535), Rural Development Administration, Republic of Korea.

In vitro maturation and in vitro fertilization of bovine oocytes cultured in a chemically defined, protein-free medium: effects of carbohydrates and amino acids

1999 ◽  
Vol 11 (2) ◽  
pp. 127 ◽  
Author(s):  
J. M. Lim ◽  
B. C. Lee ◽  
E. S. Lee ◽  
H. M. Chung ◽  
J. J. Ko ◽  
...  
Keyword(s):  
Amino Acids ◽  
In Vitro Fertilization ◽  
In Vitro Maturation ◽  
Essential Amino Acids ◽  
Defined Medium ◽  
Blastocyst Stage ◽  
Vitro Fertilization ◽  
Pronuclear Formation ◽  
Bovine Oocytes

This study was conducted to examine the effects of carbohydrates and amino acids on the maturation and fertilization of bovine oocytes. To evaluate the effect of each treatment without any unpredictable interference, oocytes were cultured in a simply defined medium (modified Tyrode’s medium; mT) without the addition of hormones and proteins. In Experiment 1, oocyte maturation to the metaphase-II stage was significantly (P<0.0001) enhanced after the addition of glucose (5.6 mМ), lactate (10 mМ) and/or pyruvate (0.5 mМ) to mT (37–74%) than after no addition (0%). In mT supplemented with glucose, the addition of 19 essential and non-essential amino acids (aa; 0, 0.01, 0.1, 1, 5 or 10%) did not further improve in vitro maturation (Experiment 2) or in vitro fertilization (Experiment 3) of oocytes. However, more (P<0.05) pronuclear formation after in vitro-insemination was found in oocytes matured in mT with 1% aa and glucose than in oocytes matured in mT with glucose alone (56% vs. 35%). Penetration of spermatozoa into the ooplasm was initiated at 3 h after insemination and pronuclear formation from 8 h (Experiment 4). When cultured inseminated oocytes were examined up to 192 h post insemination, a significant (P<0.05) increase in the number of 2-cell (18 v. 38%) and 8-cell embryos, (7 v. 20%) and morulae (0 v. 8%) was found after the addition of 1% aa to mT with glucose than after no addition (Experiment 5). A limited number of oocytes matured in mT with aa and glucose developed to the blastocyst stage (6%). These results indicate that exogenous carbohydrates and amino acids are prerequisites for the maturation and fertilization of bovine oocytes in vitro. Glucose alone promotes the nuclear maturation of oocytes, whereas amino acids aid the pronuclear formation of fertilized oocytes.

scholarly journals Bayesian Network Modeling of IVF Blastocyst Score Prediction

2021 ◽  
Vol 12 (45) ◽  
pp. 12-28
Author(s):  
Aslı Uyar ◽  
Yasemin Atılgan Şengül
Keyword(s):  
Embryo Transfer ◽  
Nearest Neighbor ◽  
Morphological Evolution ◽  
Blastocyst Stage ◽  
Single Embryo Transfer ◽  
Blastocyst Development ◽  
True Negative ◽  
Vitro Fertilization ◽  
Estimate Method

Embryo transfer may be performed at cleavage stage (on day 2-3) or at blastocyst stage (on day 5) in In-Vitro Fertilization (IVF) treatment. Elective single embryo transfer at blastocyst stage increases the pregnancy probability and reduces the number of multiple pregnancies. However, the extended culture of embryos in the laboratory may result in transfer cancelation if no high quality blastocyst develops by day 5. Predicting the blastocyst score of individual embryos may help physicians to decide whether or not to further culture the embryos in the laboratory. In this paper, we use Bayesian networks for predicting the blastocyst score by modeling the morphological evolution of IVF embryos. We propose a weighted nearest neighbor approach to adjust the frequency estimates in the conditional probability table. Experimental results show that the proposed method significantly increases the accuracy and reduces false positive rates in IVF data in comparison to the frequency estimate method. Our proposed model can also predict low quality blastocyst development with a 77.3% True Negative rate. Using this model can help preventing developmental failures of embryos during IVF treatment.

In vitro culture of embryos produced by in vitro maturation and fertilization of bovine follicular oocytes

1989 ◽  
Vol 1989 ◽  
pp. 14-14
Author(s):  
V. Vergos ◽  
A. Gordon ◽  
M. Gallagher ◽  
I. Gordon
Keyword(s):  
In Vitro Maturation ◽  
Present Report ◽  
Subsequent Development ◽  
Cumulus Cells ◽  
Blastocyst Stage ◽  
Fertilization In Vitro ◽  
Factors Affecting ◽  
Stage Of Development

A previous report from this laboratory dealt with the establishment of pregnancies in the early months of gestation after the non-surgical transfer of cattle embryos derived from the in vitro maturation (IVM) of primary bovine oocytes, their fertilization in vitro (IVF) and their subsequent development to the transferable stage (morula/blastocyst) using an in vivo (sheep oviduct) culture system (Lu et al.,1987). The present report deals with some factors affecting the efficiency of IVF and with the culture in vitro of zygotes to the morula/ blastocyst stage of development. Some embryos were frozen and after thawing transferred by non-surgical procedures to five recipient cattle to obtain information on their capacity to undergo further embryonic development.Primary oocytes, enclosed in cumulus cells, were recovered from vesicular follicles (2-6mm) after their dissection from the ovaries of heifers slaughtered at a local abattoir. The ovaries were brought to the laboratory within one hour of animal slaughter in medium held at 35'C.

Outcome of Repeated Cycles of In Vitro Fertilization With Blastocyst Stage Embryo Transfer.

2001 ◽  
Vol 75 (4) ◽  
pp. S16-S17
Author(s):  
B.S. Shapiro ◽  
K.S. Richter ◽  
D.C. Harris ◽  
S.T. Daneshmand
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Transfer ◽  
Blastocyst Stage ◽  
Vitro Fertilization ◽  
Stage Embryo
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