In vitro Embryo Production in Llamas (Lama glama) from In vivo Matured Oocytes with Raw Semen Processed with Androcoll-E using Defined Embryo Culture Media

2011 ◽  
Vol 47 (4) ◽  
pp. 562-567 ◽  
Author(s):  
V Trasorras ◽  
S Giuliano ◽  
G Chaves ◽  
D Neild ◽  
A Agüero ◽  
...  
2022 ◽  
Vol 8 ◽  
Author(s):  
Jordana S. Lopes ◽  
Cristina Soriano-Úbeda ◽  
Evelyne París-Oller ◽  
Sergio Navarro-Serna ◽  
Analuce Canha-Gouveia ◽  
...  

Assisted reproductive technologies play a major role in the cattle industry. An increase in the use of in vitro-derived embryos is currently being seen around the globe. But the efficiency and quality of the in vitro-derived embryos are substandard when compared to the in vivo production. Different protocols have been designed to overcome this issue, one of those being the use of reproductive fluids as supplementation to embryo culture media. In this study, in vitro-derived calves produced with reproductive fluids added to their embryo production protocol were followed for the first year of life pairwise with their in vivo control, produced by artificial insemination (AI), and their in vitro control, produced with standard supplementation in embryo production. The objective was to assess if any differences could be found in terms of growth and development as well as hematological and biochemical analytes between the different systems. All the analysed variables (physical, hematological, and biochemical) were within physiological range and very similar between calves throughout the entire experiment. However, differences were more evident between calves derived from standard in vitro production and AI. We concluded that the use of reproductive fluids as a supplementation to the embryo culture media results in calves with closer growth and development patterns to those born by AI than the use of bovine serum albumin as supplementation.


2010 ◽  
Vol 22 (1) ◽  
pp. 32 ◽  
Author(s):  
R. L. Krisher ◽  
M. B. Wheeler

Mammalian embryo development is still relatively inefficient in vitro. Much research has been conducted on the chemical environment, or culture medium, surrounding the embryo, but little attention has been given to the actual physical culture environment, which has changed very little over the years. The application of microfluidics to embryo production in vitro is a tantalising approach that may alleviate some of the limits that traditional microdrop culture places on embryo development and research into gamete and embryo physiology. These devices may lead to enhanced in vitro embryo development and quality by more closely mimicking the in vivo environment. Initial work in this area is promising and gives us proof-of-principle that these unique microfluidic systems may indeed be applicable to in vitro culture of gametes and embryos. The present paper reviews the advantages of microfluidics for in vitro embryo production: how the platforms are manufactured, the current uses of microfluidics in assisted reproduction, static v. dynamic culture environments, individual gamete and embryo culture and the future directions of microfluidic application to in vitro embryo production and manipulation. Finally, preliminary data from our laboratory using a new microfluidic well insert for porcine, bovine and murine embryo culture is discussed.


Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 2135
Author(s):  
Martina Colombo ◽  
Isa Mohammed Alkali ◽  
Sylwia Prochowska ◽  
Gaia Cecilia Luvoni

In vitro embryo production in cats and dogs still presents some challenges, and it needs to be optimized to transfer efficient protocols to related wild, endangered species. While the chemical composition of culture media has been the focus of several studies, the importance of culture substrates for oocyte and embryo culture has often been neglected. Traditional in vitro systems, i.e., two-dimensional cultures, do not resemble the physiological environments where cells develop, and they may cause morphological and functional alterations to oocytes and embryos. More modern three-dimensional and microfluidic culture system better mimic the structure and the stimuli found in in vivo conditions, and they could better support the development of oocytes and embryos in vitro, as well as the maintenance of more physiological behaviors. This review describes the different culture systems tested for domestic carnivore reproductive cells along the years, and it summarizes their effects on cultured cells with the purpose of analyzing innovative options to improve in vitro embryo production outcomes.


2020 ◽  
Author(s):  
Mehdi Hajian ◽  
Farnoosh Jafarpour ◽  
Sayed Morteza Aghamiri ◽  
Shiva Rouhollahi Varnosfaderani ◽  
Mohsen Rahimi ◽  
...  

Abstract Background: The ingredients of embryo culture media developed by different companies are disclosed. Thus, it is impossible to determine which ingredients might be responsible for differences in pre-and post-implantation embryo development. To address this gap, we performed an experiment to compare two embryo culture media, namely, SOF and commercial BO, on pre- and post-implantation development of cloned Sannen goat embryos. Cumulus oocyte complexes derived from slaughterhouse ovaries were used for in vitro embryo production . In vitro development of IVF, parthenogenetic and SCNT embryos were assessed in both BO and SOF media. The expression of 16 genes, including AKT , OCT4 , SOX2 , BMPR1 , FGFR4 , CDC25 , CDX2 , GCN5 , PCAF , FOXD3 , SMAD5 , FZD , LIFR1 , CTNNB , ERK1 , and IFNT , belonging to 7 important pathways, i.e. pluripotency, FGF, TGFβ, cell cycle and proliferation, histone transferase, trophectoderm, and WNT, were examined in the goat SCNT and IVF blastocysts from both BO and SOF media. Results: The blastocyst rate in BO medium was significantly higher than that of the SOF medium in SCNT embryos ( P < 0.05). All of the genes examined showed increased expression levels in SCNT embryos compared to IVF embryos. In the IVF group, OCT4 , BMPR1 , and GCN5 showed significantly higher expression in the SOF medium compared to the BO medium. In this group, AKT , FGFR4 , SOX2 showed significantly lower expression in the SOF medium compared to the BO medium. In the SCNT group, FGFR4 , GCN5 , FZD , CTNNB , BMPR1 , and FGFR4 showed significantly higher expression in SOF medium compared to BO medium. In vivo development did not differ significantly between the two groups. Conclusions: Based on these results, we concluded that the limited information available on the allocations of ICM and TE cells in SCNT embryos and embryo-specific gene expression may be the major drawback IVC medium and an impediment to successful animal cloning.


1997 ◽  
Vol 47 (1) ◽  
pp. 259 ◽  
Author(s):  
L.M.T.E. Lansbergen ◽  
E.H.A.T. Hanenberg ◽  
A.M. van Wagtendonk-de Leeuw

2004 ◽  
Vol 16 (2) ◽  
pp. 259
Author(s):  
J.R. Miles ◽  
C.E. Farin ◽  
K.F. Rodriguez ◽  
J.E. Alexander ◽  
P.W. Farin

The role of the vascular supply in the development of placentas from embryos produced in vitro is poorly understood. The objective of this study was to determine the effects of in vitro embryo production on morphometry of blood vessels within fetal (cotyledonary) and maternal (caruncular) components of the placentome during late gestation. In vivo-produced embryos were recovered from superovulated Holstein cows on Day 7 after estrus. For in vitro embryo production, oocytes were aspirated from the ovaries of Holstein cows, matured in vitro, and then fertilized. Presumptive zygotes with their cumulus cells were transferred into M-199 with 10% estrus cow serum and cultured for 168h post-insemination. Semen from the same Holstein sire was used for the production of in vivo and in vitro embryos. Single blastocysts from each production system were transferred into the uteri of heifers. On Day 222 of gestation, fetuses and placentas were recovered in utero (in vivo, n=12; in vitro, n=12). Placentomes were collected, fixed and sectioned. Fetal and maternal blood vessels were identified within placentome sections using immunocytochemistry for vascular endothelial growth factor (VEGF) protein. A total of 4.8×105μm2 of tissue were examined from each placentome. Stereological methods were used to determine the volume densities of fetal and maternal blood vessels. Data were analyzed by GLM procedures. Fetuses were heavier (P=0.03) in the in vitro group (20.7±1.0kg, LS mean±SEM) compared to the in vivo group (17.3±1.0kg). Placentas were also heavier (P=0.06) for the in vitro group (2.5±0.2kg) compared to the in vivo group (2.0±0.2kg). Placental efficiency, calculated as fetal weight/placental weight, was similar between the two treatment groups (9.0±0.5 and 8.9±0.5 for in vivo and in vitro, respectively). Fetal vascular volume density in placentomes was not different between the two treatment groups (5.4±0.3% and 5.4±0.3% for in vivo and in vitro, respectively). In contrast, maternal vascular volume density was greater (P=0.02) for placentomes in the in vitro group (5.9±0.3%) compared to in vivo controls (4.9±0.3%). In summary, compared to placentomes from embryos produced in vivo, placentomes from embryos produced in vitro had similar volume density of fetal vessels, but had significantly increased volume density of maternal vessels. Supported by the State of North Carolina.


2014 ◽  
Vol 26 (1) ◽  
pp. 162
Author(s):  
H. Tribulo ◽  
J. Carcedo ◽  
R. Tribulo ◽  
J. Menajovsky ◽  
B. Bernal ◽  
...  

An experiment was designed to evaluate in vivo and in vitro embryo production following the use of frozen–thawed conventional or Y-sexed semen from a Brangus bull with known high fertility. For in vivo embryo production, Brangus heifers (n = 12) were superovulated twice in a crossover design and inseminated with sexed or conventional semen. On Day 0, all heifers received an intravaginal progesterone device (DIB 1 g, Syntex S.A., Buenos Aires, Argentina) and 2.5 mg oestradiol benzoate and 50 mg progesterone (Progestar, Syntex S.A.) by intramuscular injection (IM). On Day 4, heifers were superstimulated with 200 mg of NIH-FSH-P1 Folltropin-V (Bioniche Animal Health, Belleville, Ontario, Canada) in twice-daily decreasing doses over 4 days. In the a.m. and p.m. of Day 6, all heifers received PGF2a (Ciclase, Syntex) and DIBs were removed in the p.m.. In the a.m. of Day 8, heifers received 100 μg de Gonadolerin (Gonasyn, Syntex S.A.) and were randomly allocated to receive either one straw of conventional semen (24 × 106 sperm per dose) 12 and 24 h later or two straws of sexed semen (2.4 × 106 sperm per dose) 18 and 24 h after GnRH. Ova/embryos were collected nonsurgically on Day 15 and evaluated following IETS recommendations. Means were compared by t-test. Mean ( ± s.e.m.) number of ova/embryos, fertilized ova, and transferable embryos were 14.8 ± 2.7, 9.4 ± 1.8, and 7.1 ± 1.7 v. 16.8 ± 3.1, 9.9 ± 2.5, and 8.1 ± 2.0 for donors inseminated with conventional or sexed semen, respectively (P > 0.6). For in vitro production, oocytes were obtained from 50 ultrasound-guided follicle aspiration (OPU) sessions that was performed at random stages of the oestrous cycle and without superstimulation in 22 Brangus cows and heifers. Oocytes were classified and matured in TCM-199 medium with NaHCO3 and supplemented with 1% fetal bovine serum. Semen samples from the same bull used for in vivo embryo production were selected using Percoll and capacitated in Fert medium and used at a final concentration of sperm/mL for nonsexed semen and 2 × 106 sperm mL–1 for sexed semen. After 16 h (sexed) or 18 h (conventional) in Fert medium, zygotes were denuded and cultured in SOF supplemented with 0.4% BSA under oil at 37°C, 5% CO2 and saturated humidity for 7 days. The total number of oocytes matured and fertilized was 528 and 318 for conventional and sexed semen, respectively. Means were compared by t-test and proportions by chi-squared test. Mean (± s.e.m.) number of cleaved zygotes and blastocysts produced per OPU session did not differ between conventional (11.0 ± 1.4 and 7.1 ± 1.0) and sexed (8.7 ± 0.8 and 4.9 ± 0.7; P > 0.2) semen. However, the proportion of cleaved zygotes and blastocysts produced were significantly higher (P < 0.05) with conventional semen (61.2%; 329/538 and 39.4%; 212/538) than with sexed semen (54.4%; 173/318 and 30.8%; 98/318), respectively. In conclusion, comparable number of embryos can be obtained in vivo with sexed or conventional semen from a bull with proven high fertility. However, the proportion of blastocysts produced in vitro is likely to be reduced following the use of sexed as compared with conventional semen from the same bull.


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