In vitro Production of Bovine Blastocysts by in vitro Maturation and Fertilization of Oocytes and Subsequent in vitro Culture

1989 ◽  
Vol 24 (3) ◽  
pp. 134-139 ◽  
Author(s):  
U. Berg ◽  
G. Brem
Keyword(s):  
In Vitro Culture ◽  
In Vitro Maturation ◽  
In Vitro Production ◽  
Vitro Production

scholarly journals Use of Melatonin in the In Vitro Production of Bovine Embryos

2020 ◽  
Vol 21 ◽  
Author(s):  
Alan da Silva LIRA ◽  
Ricardo de Macedo CHAVES ◽  
Felipe de Jesus MORAES JUNIOR ◽  
Sergio Henrique COSTA JUNIOR ◽  
Brenda Karine Lima do AMARAL ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Control Group ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Maturation Medium ◽  
Significant Difference ◽  
Maturation Rate ◽  
Vitro Production

ABSTRACT We aimed to assess the effects of melatonin in the in vitro production of bovine embryos. Our experiment was conducted at the Laboratório de Reprodução Animal of the Universidade Estadual do Maranhão. The cumulus-oocyte complexes (COCs) were distributed among treatments at concentrations of 0, 10-1, 10-3 and 10-5 µMol/L melatonin. Our experiment was further divided into two: the first was to assess the effect of different concentrations of melatonin (treatments) on the maturation rate of COCs, and the second was to assess the effects of melatonin treatments on the in vitro production of bovine embryos. The results from the first experiment demonstrated no significant difference between the in vitro maturation rate of the cultivated COCs in treatments with melatonin. In the second experiment, however, melatonin treatments yielded statistically higher cleavage, morula and blastocyst rates in the 10-5 µM group (52.9%, 52.9%, and 35.3%, respectively), and lower rates in the 10-1 µM group (19.5%, 19.5% and 7.8%, respectively), compared to the others. The control group (no melatonin) and the 10-3 µM group showed similar results. We concluded that supplementation of melatonin in the in vitro maturation medium resulted in no improvement in the oocyte maturation rate, but in the in vitro production of embryos at different concentrations, the 10-5 µM group displayed better results, but with no improvement in the variables (P < 0.05).

scholarly journals Effects of in vitro maturation media on in vitro fertility of porcine oocytes and early development of embryos

2020 ◽  
Vol 18 (2) ◽  
pp. 249-255
Author(s):  
Nguyen Viet Linh ◽  
Nguyen Thi Hiep
Keyword(s):  
Follicular Fluid ◽  
In Vitro Maturation ◽  
Subsequent Development ◽  
Developmental Competence ◽  
In Vitro Production ◽  
Vitro Fertilization ◽  
Fetal Bovine ◽  
Vitro Production ◽  
Normal Fertilization

In pigs, embryo productivity is still lower than that in other livestocks. One of the reasons is incomplete maturation of porcine oocytes in in vitro conditions. Therefore in vitro maturation (IVM) plays a crucial role in in vitro production of porcine embryos. It provides prerequisite condition to in fertilization and subsequent development of porcine embryos. In a previous study, effects of NCSU-37-based medium and TCM-199-based media supplemented with porcine follicular fluid (pFF) or Fetal Bovine Serum (FBS) on in vitro maturation of Landrace oocytes collected in Vietnam have been compared, suggesting that NCSU-37 medium supplemented with 10% of porcine follicular fluid (pFF) had the highest rate of oocytes reach to metaphase II stage in comparison to those of the other two TCM-199-based media. In the present study, further experiments were carried out to evaluate the contribution of IVM media on fertilization capability and developmental competence. Porcine oocytes matured in vitro in 3 media: NCSU-37 supplemented with 10% pFF, TCM-199 supplemented with either 10% pFF or 10% FBS were subjected to in vitro fertilization and subsequent in vitro culture to monitor fertility and embryo development. The results showed that penetration and normal fertilization rates in both TCM-199 groups are both higher than that of NCSU-37 group. Moreover, the cleavage and blastocyst rates, and cell numbers of blastocysts which is a criterion for embryo quality were all higher in TCM-199 groups, especially in the group supplemented with pFF. It might be concluded that TCM-199 media supplemented with either pFF or FBS are suitable for effective in vitro maturation of Landrace porcine oocytes collected in Vietnam.

272 A NEW APPROACH TO IN VITRO MATURATION (IVM) AND EMBRYO IN VITRO PRODUCTION: INDUCED IVM SUBSTANTIALLY IMPROVES EMBRYO YIELD AND PREGNANCY OUTCOMES

2010 ◽  
Vol 22 (1) ◽  
pp. 293
Author(s):  
R. B. Gilchrist ◽  
F. K. Albuz ◽  
J. G. Thompson
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Cumulus Cell ◽  
In Vitro Production ◽  
New Approach ◽  
Fetal Survival ◽  
Wide Range ◽  
Vitro Production

Oocyte in vitro maturation (IVM) is the rate-limiting step in the in vitro production (IVP) of embryos. Oocyte maturation in vivo is a highly orchestrated, induced process, whereby cAMP-mediated meiotic arrest is overridden by the gonadotrophin surge prior to ovulation. However, aspirated oocytes resume maturation spontaneously compromising developmental competence. Hence, we hypothesized that establishing an induced system in vitro would synchronize oocyte-somatic cell communication leading to improved oocyte quality. Abattoir-collected bovine or 129/Sv mouse oocytes were treated for the first 1 to 2 h in vitro (pre-IVM) with the adenylate cyclase activator forskolin (100 μM, 50 μM, respectively) and a nonspecific phosphodiesterase (PDE) inhibitor, IBMX (500 μM, 50 μM), which substantially increased cumulus-oocyte complex (COC) cAMP (bovine, 180 v. 2 fmol/COC, treated v. control; P < 0.001). To maintain oocyte cAMP levels and prevent precocious oocyte maturation, IVM media (VitroMat + BSA) contained an oocyte-specific (type 3) PDE inhibitor, cilostamide (20 μM, 0.1 μM), plus FSH to induce maturation. The net effect of this system (induced-IVM) was to increase oocyte-cumulus cell gap-junctional communication (bovine: 1000 ± 148 v. 340 ± 73 unit, treated v. control; P < 0.05) and to slow meiotic progression through prophase I to metaphase II, extending the normal IVM interval (bovine: 30 v. 24 h, mouse: 22 v. 18 h; treated v. control). FSH was required to complete maturation and FSH-induced maturation was prevented by an epidermal growth factor receptor inhibitor, AG1478 (2.5 μM), demonstrating induced oocyte maturation functions via secondary autocrine signaling within the cumulus cell compartment. These effects on COC functions had profound consequences for oocyte developmental potential. In completely serum-free bovine IVP, induced-IVM more than doubled blastocyst yield (69 v. 27%, treated v. control; P < 0.05) and improved blastocyst quality (186 v. 132 blastomeres). To achieve these rates, the pre-IVM phase, the modified IVM conditions, and delayed IVF were all required. Adapting the system to the mouse, induced-IVM increased blastocyst rate (86 v. 55%, treated v. control; P < 0.05), implantation rate (51 v. 25%; P < 0.01), fetal survival rate (29 v. 5%; P < 0.01) and fetal weight (0.9 v. 0.5 g; P < 0.01). All these embryonic and fetal outcomes in mice were equivalent (P > 0.05) using induced-IVM to levels obtained from in vivo-matured control oocytes (conventional IVF). Data were analyzed by ANOVA. In conclusion, induced-IVM mimics some of the characteristics of oocyte maturation in vivo and substantially improves oocyte developmental outcomes in 2 disparate mammalian species. Adaption of this new approach to clinical/field conditions should lead to new opportunities for a wide range of reproductive biotechnologies. Such a notable increase in IVM efficiency could see IVP as the preferred embryo production technology in future livestock artificial breeding programs. Funded by an Australian Research Council Linkage Grant and Cook Australia. Thanks to M. Sasseville, M. Lane, and D. T. Armstrong.

scholarly journals Collection grading and evaluation of cumulus-oocyte-complexes for in vitro maturation in sheep

1970 ◽  
Vol 28 (1) ◽  
pp. 31-38
Author(s):  
MNS Talukder ◽  
A Iqbal ◽  
MAMY Khandoker ◽  
MZ Alam
Keyword(s):  
Corpus Luteum ◽  
Culture Condition ◽  
In Vitro Maturation ◽  
Type I ◽  
Type Ii ◽  
In Vitro Production ◽  
Length Width ◽  
Left And Right ◽  
Vitro Production

An alternative to superovulation is in-vitro production (IVP) of embryos where the efficient collection and grading of oocytes is important. Ovaries from an abattoir were collected and categorized as type I with no corpus luteum (CL), and type II with CL. The length, width and weight of type I and type II ovary were 1.4 ± 0.03 and 1.5 ± 0.08 cm; 0.8 ± 0.04 and 1.0 ± 0.07 cm; 0.6 ± 0.07 and 0.7 ± 0.04 gm, respectively, each significantly (P<0.05) higher in type II ovaries. A total of 80 and 78 follicles were observed and 60 and 61 follicles aspirated from left and right ovaries, respectively, from each of 25 ovaries. Out of 133 follicles 100 were aspirated from 40 type-I ovaries, and 21 aspirated from 10 type-II ovaries. The differences in the number of normal, abnormal and total cumulus-oocytecomplex (COCs) per ovary between left and right ovaries were not significant (P>0.05). The number of normal (1.9 ± 0.11) and total (2.5 ± 0.14) COCs per ovary were significantly (P<0.05) higher in ovaries without than in those with CL (1.2 ± 0.36 and 2.0 ± 0.30, respectively). But the number (0.80 ± 0.13) of abnormal COCs per ovary was significantly (P<0.05) higher in ovaries with CL than in those without (0.7 ± 0.09). Significantly (P<0.05) higher percentage of COCs expansion was grade A (6.9 ± 2.05) than grade B (53.1 ± 1.27) COCs. It is suggested that type I (without CL) ovaries and follicles of 2-6 mm diameter are suitable to collect good quality COCs for in-vitro maturation (IVM) of oocytes and the culture condition for IVM of sheep COCs are reported. DOI: http://dx.doi.org/10.3329/bvet.v28i1.8811   Bangl. vet. 2011. Vol. 28, No. 1, 31-38

Laboratory production of buffalo (Bubalus bubalis) embryos

1998 ◽  
Vol 10 (5) ◽  
pp. 379 ◽  
Author(s):  
P. Palta ◽  
M. S. Chauhan
Keyword(s):  
In Vitro Maturation ◽  
Large Scale ◽  
Follicle Stimulating Hormone ◽  
Bubalus Bubalis ◽  
Blastocyst Stage ◽  
In Vitro Production ◽  
Nuclear Maturation ◽  
Fertilization Rates ◽  
Vitro Production

There is an increasing interest in large-scale in vitro production (IVP) of buffalo embryos through in vitro maturation (IVM), fertilization (IVF) and culture (IVC) of oocytes for faster multiplication of superior germplasm. The recovery of total and usable quality oocytes from slaughterhouse ovaries is low in this species. The nuclear maturation rates of buffalo oocytes matured in the presence of follicular fluid or serum and hormones like luteinizing hormone, follicle-stimulating hormone and oestradiol vary from 70 to 80% and are comparable to those reported for cattle oocytes. However, with fertilization rates of 40–55%, and the yield of blastocysts at around 10–15%, the efficiency of IVP is much lower than that in cattle. The in vitro sperm preparation procedures and the systems employed for performing IVF and culture of zygotes up to blastocyst stage are suboptimal and need substantial improvements. The quality and viability of blastocysts produced need to be checked by cell count, and after transfer to synchronized recipients, for development of quality control standards.

scholarly journals Supplementation of in vitro maturation medium with serum from dairy cows at early and late lactation in the in vitro production of bovine embryos

2018 ◽  
Vol 12 (4) ◽  
pp. 311
Author(s):  
Letícia Franco Collares ◽  
Jorgea Pradieé ◽  
Morgana Alves Borges ◽  
Bruna Mion ◽  
Patrícia Gindri ◽  
...  
Keyword(s):  
Dairy Cows ◽  
In Vitro Maturation ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Maturation Medium ◽  
Vitro Production

158 EFFECTS OF APOTRANSFERRIN ON IN VITRO MATURATION OF CUMULUS - OOCYTES COMPLEXES (COCs) IN HANWOO, KOREAN NATIVE COWS

2006 ◽  
Vol 18 (2) ◽  
pp. 187
Author(s):  
S.-H. Choi ◽  
S.-R. Cho ◽  
M.-H. Han ◽  
H.-J. Kim ◽  
C.-Y. Choe ◽  
...  
Keyword(s):  
In Vitro Maturation ◽  
Blastocyst Stage ◽  
Basic Fuchsin ◽  
In Vitro Production ◽  
Control Groups ◽  
Significant Difference ◽  
Native Cattle ◽  
Vitro Development ◽  
Vitro Production

For in vitro production of embryos, animal sera have been used as energy sources, maturation promoters, vitamins, growth factors, and antioxidative compounds. However, the sera had risk of virus and mycoplasma infections which could result in too big offspring and cause dystocia in ovine and bovine. Apotransferrin (apo-Tf) is a component of mammalian sera and has played a role as an antioxidant in media. A study was conducted to investigate the effects of apo-Tf on in vitro maturation of cumulus-oocytes complexes (COCs) in Hanwoo, Korean native cows. Ovaries were collected from a slaughterhouse and COCs were taken from 2-6-mm antral follicles. The collected COCs were washed three times with 0.1M polyvinyl alcohol (PVA)-TCM199 and matured in 0, 1, 10, or 100 �g/mL apo-Tf with TCM-199 at 39�C, 5% CO2, 95% air for 6, 12, or 24 h. Mature COCs were fertilized with frozen-thawed Korean native cattle semen treated with BO medium (Brackett and Oliphants 1975 Biol. Reprod. 12, 260-274) containing 5 mM caffeine and 1 �g/mL heparin for 8 h and developed to the blastocyst stage in 5% FBS and 0.3% BSA in TCM199-IVMD (IFP, Japan). To evaluate the morphology of nuclear types, the matured COCs were fixed in 1:3 acetic acid-ethanol for 30 s and stained with 3% basic Fuchsin. IVM and IVF were replicated three times. All of the results were analyzed by ANOVA using the STATVIEW program. The maturation rates of control were 34.2%, 37.3%, and 45.8% for 6, 12, and 24 h, respectively. There were no differences among the concentrations of apo-Tf, and nuclear types at 78.3-87.0% GVBD for 6 h, 82.8-91.3% MI for 12 h, and 88.9-100.0% MII for 24 h, with 1, 10, and 100 �g/mL apo-Tf, respectively. Conversely, there was significant difference between 1 �g/mL and 10 �g/mL in terms of cleavage rates, although the others did not vary significantly (P < 0.05). There were significant differences among the concentrations of apo-Tf for blastocyst formation (P < 0.05). Blastocysts matured with 1, 10, and 100 �g/mL apo-TF and developed in 5% FBS and 0.3% BSA in TCM199-IVMD showed rates of 8.8-21.6%, 9.4-35.3%, and 9.1-19.1%, respectively. The control groups developed to the blastocyst stage showed rates of 8.6%, 10.8%, and 10.5% in 5% FBS and 0.3% BSA in TCM199-IVMD, respectively. These results suggest that apo-Tf is an important factor for the in vitro maturation and in vitro development of bovine COCs.

scholarly journals Meiotic maturation and in vitro maturation of bovine oocytes

2006 ◽  
Vol 22 (1-2) ◽  
pp. 29-34 ◽  
Author(s):  
Tatjana Smiljakovic ◽  
Wolfgang Tomek
Keyword(s):  
Meiotic Division ◽  
In Vitro Maturation ◽  
Meiotic Maturation ◽  
In Vitro Production ◽  
Vitro Production ◽  
Bovine Oocytes

In vitro production of embryos, and, as part of this method, in vitro maturation of oocytes, have received great attention in last ten-fifteen years. It is well established in bovine. Here, in this review is presented importance of this method, usual meiotic division is described, as well, as importance of biochemical investigations of several protein factors and enzymes, which control these processes.

scholarly journals VOLUME DE MEIO E NÚMERO DE OÓCITOS PARA FECUNDACÃO NA PRODUÇÃO IN VITRO DE EMBRIÕES BOVINOS

2004 ◽  
Vol 9 (2) ◽  
Author(s):  
D.S. BRUM ◽  
F.G. LEIVAS ◽  
M.R. NOEBAUER ◽  
M.L. BERNARDI ◽  
C.A.M. SILVA ◽  
...  
Keyword(s):  
In Vitro Maturation ◽  
Hatching Rate ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Oviduct Fluid ◽  
Vitro Production ◽  
Hatching Rates

A origem dos Complexos Cumulus-Oócitos (CCO) bovinos e os diferentes protocolos entre equipes geram variações nos índices de produção in vitro (PIV). O processo pode ser influenciado pelo número de CCO e volume das gotas de maturação, fecundação e cultivo. Neste estudo, avaliou-se a densidade de CCO e o volume de meio de fecundação sobre a PIV com 672 oócitos bovinos. A maturação in vitro (MIV) foi conduzida com 20 CCO em 200µL de TCM-199+rFSHh+10% de soro de vaca em estro (SVE), por 24h, em estufa a 39ºC, 5% CO2 em ar e umidade saturada. Na fecundação (Dia 0 =D0) utilizou-se 5, 10 ou 20 CCO e 2 volumes de meio (1:5 e 1:10), num fatorial 3x2, em 6 repetições. O sêmen foi selecionado por migração ascendente e para a fertilização (1x106 espermatozóides/mL) os gametas foram incubados em Fert-Talp, por 18h. O cultivo foi em grupos de 20 zigotos, em 200µL de fluído sintético do oviduto (SOF) aaci+5%SVE, por 8 dias e as avaliações em D2 (clivagem), D7 (Blastocistos), D9 (Blastocistos expandidos, em eclosão e eclodidos). As taxas de clivagem (80, 85 e 87%), embriões em D9 (23, 27 e 28%) e de eclosão (42, 37 e 48%) foram semelhantes (P>0,05) para 5, 10 ou 20 CCO, respectivamente. Não houve diferença (P>0,05) na clivagem (83 e 85%), embriões em D9 (24 e 28%) e eclosão (41 e 44%) entre as proporções 1:5 e 1:10µL. A produção de blastocistos e taxa de eclosão em D9 não são influenciados pelo número de CCO, nem pela proporção de CCO por volume de meio na fecundação. Volume of medium and number of oocytes for fertilization in the in vitro production of bovine embryos Abstract The origin of the bovine Cumulus Oocytes Complexes (COC) and the different protocols among teems are responsible for the variation rates of the in vitro production (IVP). The process is also influenced by COC number and by the volume of the drop in the maturation, fertilization and culture. In this study, the effect of bovine COC density per drop medium and the fertilization medium volume were evaluated during the bovine IVP using 672 COC. The in vitro maturation was performed with 20 COC in 200µL of TCM-199+rFSHh+10% estrus cow serum (ECS), for 24h in an incubator with 5%CO2 in air, saturated humidity, at 39oC. For the fertilization (Day 0 = D0) 5, 10 or 20 COC and 2 volumes ratio of medium (1:5 and 1:10) were used in a factorial outline 3x2, in 6 replications. Thawed semen was prepared by a swim up process and for the fertilization (1x106 spermatozoa/ ml) the gametes were maintained in Fert-Talp for 18h. Groups of twenty presumed zygotes were cultured in 200µL synthetic oviduct fluid (SOFaaci)+5% ECS, for 8 days. The evaluations were carried out in D2 (cleavage), D7 (blastocysts) and D9 (expanded and hatched blastocyts). The rates of cleavage (80, 85 and 87%), embryos in D9/ D2 (23, 27 and 28%) and hatched blastocstys in D9/D7 (42, 37 and 48%) were similar (P>0.05) for 5, 10 and 20 COC, respectively. No difference was observed in cleavage (83 and 85%), embryos in D9 (24 and 28%) and hatching rates (41 and 44%) for 1:5 and 1:10µL of fertilization medium. Blastocysts production and the hatching rate in D9 were neither influenced by the COC number, nor by the proportion of COC per medium volume used during fertilization.

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