High developmental potential in vitro and in vivo of cattle embryos cloned without micromanipulators

Rodríguez, Lleretny; Navarrete, Felipe; Tovar, Heribelt; Cox, J.F.; Castro, Fidel Ovídio

doi:10.1007/s10815-007-9194-x

Cited by 19 publications

(14 citation statements)

References 27 publications

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“…Conventional production of elongated embryos in vivo followed by non-surgical flushing or flushing from collected uteri produced similar results to ours (Degrelle et al, 2005;Arnold et al, 2006;Sawai et al, 2007;Rodríguez et al, 2008). Nonetheless elongated bovine embryos had been produced both in vivo and in vitro.…”

Section: Elongation Of Bovine Embryos In Heterologous and Homologous supporting

confidence: 81%

Elongation and gene expression in bovine cloned embryos transferred to temporary recipients

Rodríguez-Álvarez

Cox

Navarrete

et al. 2009

Zygote

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SummaryElongated embryos provide a unique source of information about trophoblastic differentiation, gene expression and maternal-embryonic interactions; however they are difficult and costly to obtain, especially elongated cloned embryos. One alternative is their production in heterologous temporary recipients such as sheep and goats. We aimed to produce elongated bovine cloned embryos using heterologous transfer to temporary recipients. Day-7 cloned cattle blastocysts were transferred to the uteri of ewes and goats and recovered as elongated structures at day 17. We evaluated elongation, length, presence of embryonic disc and expression of several important genes for embryonic development. We also produced homologous (cloned cattle embryos transferred into cattle uteri). Cloned bovine blastocysts were able to proceed with preimplantation development through elongation with high efficiency despite the species to which they were transferred. In qualitative and quantitative RT-PCR experiments we found differences in the pattern of gene expression among embryos recovered from different species. Sox2, Nanog and FGF-4 were markedly deregulated. No previous reports about the expression pattern of the studied genes had been published for elongated bovine cloned embryos produced in intermediate recipients, furthermore, the pattern of expression of Nanog, Oct4, Eomes, Cdx2, IFN-tau, Dicer, FGF-4 and Sox2 shown here are novel for elongated cloned bovine embryos created by hand-made cloning. Our data confirmed that sheep and goats can be used as temporary recipients. This model could serve as a basis for further research on gene expression and cellular changes during bovine peri-implantation development.

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Section: Elongation Of Bovine Embryos In Heterologous and Homologous supporting

confidence: 81%

Elongation and gene expression in bovine cloned embryos transferred to temporary recipients

Rodríguez-Álvarez

Cox

Navarrete

et al. 2009

Zygote

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“…COCs were obtained from the ovaries of two abattoirs (Frigosur Ltda, Chillan, VIII Region–Bio‐Bio and Matadero Angol, IX Region‐Araucanía) following the standard procedure described by Rodríguez, Navarrete, Tovar, Cox, and Castro (). Animal breed and productivity was identified at the moment of ovary collection and grouped by dairy cattle (more than two lactations; Holstein–Friesian breed) or beef cattle.…”

Section: Methodsmentioning

confidence: 99%

Effect of BMP15 and/or AMH during in vitro maturation of oocytes from involuntarily culled dairy cows

Velásquez

Mellisho

Castro

et al. 2018

Molecular Reproduction Devel

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The high metabolic activity to which the dairy cattle are exposed to maintain milk production altered steroid metabolism that affects reproductive physiology and reduce oocyte competence. Our aims were (a) to characterize the competence of immature oocytes collected from dairy cattle based on the expression of genes in cumulus cells (CCs) and (b) to improve oocyte competence to support preimplantation embryo development by the supplementation of maturation medium with bone morphogenetic protein 15 (BMP15) and/or anti‐mullerian hormone (AMH). Oocyte donors were identified at the moment of ovary collection and grouped by involuntarily culled dairy cows (Holstein breed) or beef cattle. The embryo development speed to blastocyst of the cull dairy cattle versus beef cattle (control group) was lower. Besides, <10% of oocytes (with CC biopsies) derived from dairy cattle were able to develop to the blastocyst stage. In addition, a higher level of expression and a positive correlation were observed in the expression of most of the genes evaluated (LUM, KRT18, KRT8, CLIC3, BMPR1B, and SLC38A3) in the cumulus–oocyte complexes that produced blastocysts versus those which did not develop correctly (arrested development). Further, use of BMP15 in the maturation of oocytes from dairy cattle seems to increase competence, modulating the expression of OCT4, SOX2, CDX2, GATA6, and TP1 in resulting blastocysts.

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“…Ovaries from beef cattle were collected at an abattoir house Frigosur Ltda, Chillan, VIII Region -Bio-Bio located 8 km (15min drive) from the laboratory, following standard procedures described by Rodríguez et al (2008). The cumulus-oocyte complexes (COCs) were in vitro matured (IVM) in four-well dishes (25-30 COCs per 500 µL well) in maturation media for 20-22 h as described (Velasquez et al 2016).…”

Section: In Vitro Embryo Productionmentioning

confidence: 99%

Extracellular vesicles secreted during blastulation show viability of bovine embryos

et al. 2019

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Extracellular vesicles (EVs) secreted by blastocysts may be clinically relevant, as indicator of embryo viability on in vitro fertilization. We tested if the characteristics of EVs secreted during blastulation are related to embryo viability. Morulae were individually cultured in SOF media depleted of EVs until day 7.5 post IVF. Viable embryos were determined by a system of extended in vitro culture of bovine embryos until day 11 (post-hatching development). Afterward, a retrospective classification of blastocyst and culture media was performed based on blastulation time (early blastulation (EB) or late blastulation (LB)) and post-hatching development at day 11 (viable (V) or non-viable embryo (NV)). A total of 254 blastocysts and their culture media were classified in four groups (V-EB, NV-EB, V-LB, NV-LB). Group V-EB had a larger blastocyst diameter (170.8 μm), higher proportion of good-quality blastocysts (77%) and larger mean size of population of EVs (122.9 nm), although the highest concentration of EVs (5.75 × 109 particles/mL) were in group NV-EB. Furthermore, small RNA sequencing detected two biotypes, miRNA (86–91%) and snoRNA (9–14%), with a total of 182 and 32 respectively. In differential expression analysis of miRNAs between V versus NV blastocysts, there were 12 miRNAs upregulated and 15 miRNAs downregulated. Binary logistic regression was used to construct a non-invasive novel model to select viable embryos, based on a combination of variables of blastocyst morphokinetics and EVs characteristics, the ROC-AUC was 0.853. We concluded that characteristics of EVs secreted during blastulation vary depending on embryo quality.

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High developmental potential in vitro and in vivo of cattle embryos cloned without micromanipulators

Cited by 19 publications

References 27 publications

Elongation and gene expression in bovine cloned embryos transferred to temporary recipients

Elongation and gene expression in bovine cloned embryos transferred to temporary recipients

Effect of BMP15 and/or AMH during in vitro maturation of oocytes from involuntarily culled dairy cows

Extracellular vesicles secreted during blastulation show viability of bovine embryos

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