Production of dromedary (Camelus dromedarius) embryos by IVM and IVF and co-culture with oviductal or granulosa cells

Khatir, H.; Anouassi, A.; Tibary, Ahmed

doi:10.1016/j.theriogenology.2004.01.016

Cited by 49 publications

(55 citation statements)

References 24 publications

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“…Trasorras et al (2012) found a lower rate of blastocysts reaching 9%.Recently, blastocyst rate ranged between 34-36% using TALP culture medium in the same living species (Trasorras et al 2014). As a result of this, cleavage rate of camel oocytes varies according to the type of culture medium (Khatir et al 2005), the size of follicle (Khatir et al 2007) and co-culture with granulosa or oviductal cells (Khatir et al 2004).…”

Section: Discussionmentioning

confidence: 99%

In vitro production of Sudanese camel (Camelus dromedarius) embryos from epididymal spermatozoa and follicular oocytes of slaughtered animals

Abdel-Khalek¹,

Gabr²,

Khalil³

et al. 2017

Polish Journal of Veterinary Sciences

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Application of assisted reproductive technology in camelidea, such as artificial insemination (AI) and embryo transfer, has been slow in comparison to that for other livestock species. In Egypt, there are few attempts to establish in vitro maturation (IVM) and fertilization (IVF) techniques in dromedary camel. The present study was carried out to produce Sudanese camel embryos using in vitro matured oocytes and epididymal spermatozoa. Dromedary camel ovaries were collected from abattoirs and then, the oocytes were aspirated from all the visible follicles on the ovarian surface (~2-8 mm in a diameter). Meanwhile, Fetal Dromedary Camel Serum (FDCS) was obtained from camel fetuses after slaughtering. Thereafter, only Cumulus Oocyte Complexes (COCs) were matured in vitro in the Tissue Culture Medium (TCM-199) complemented with 10% FDCS. Spermatozoa required for in vitro fertilization were collected from testes (epididymal cauda) of the slaughtered camel bulls. The results clearly showed that the maturation rate of oocytes at metaphase II was about 59.5% while the fertilization rate was around 70.4%. Intriguingly, the embryo rates determined were 13.1%, in 2-cell; 0.0%, in 4-cell; 34.7%, in 8-16% cell; 39.1%, in morula and 13.1% in a blastocyst stage. This study represented a successful in vitro production of Sudanese dromedary camel embryos from epididymal sperm cells and in vitro matured oocytes recovered from slaughtered camels.

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Section: Discussionmentioning

confidence: 99%

In vitro production of Sudanese camel (Camelus dromedarius) embryos from epididymal spermatozoa and follicular oocytes of slaughtered animals

Abdel-Khalek¹,

Gabr²,

Khalil³

et al. 2017

Polish Journal of Veterinary Sciences

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“…In addition, after co-incubation of equine gametes with oviduct cells, the IVF rate was not different between porcine and equine oviduct cells. The beneficial effect of homologous oviduct cells on in vitro production of embryos is also demonstrated in human [5,6], bovine [7-9], porcine [10,11], deer [12,13] and dromedary [14] species. Several studies show a positive effect of heterologous oviduct cells on gametes, fertilization and embryo development.…”

Section: Discussionmentioning

confidence: 99%

“…The secretion of oviduct secretory cells increases during the follicular phase under oestrogen and LH stimulation [3,4]. Some studies show that oviduct epithelial cells (OEC) co-culture promotes in vitro production of embryos in human [5,6], bovine[7-9], porcine [10,11], deer [12,13] and dromedary [14]. Moreover, the oviduct proteins have been shown to interact with gametes and to improve efficiency of in vitro fertilization (IVF) in porcine [10,15], bovine [9] and human [16].…”

Section: Introductionmentioning

confidence: 99%

The secretions of oviduct epithelial cells increase the equine in vitrofertilization rate: are osteopontin, atrial natriuretic peptide A and oviductin involved?

Mugnier

Kervella

Douet

et al. 2009

Reprod Biol Endocrinol

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BackgroundOviduct epithelial cells (OEC) co-culture promotes in vitro fertilization (IVF) in human, bovine and porcine species, but no data are available from equine species. Yet, despite numerous attempts, equine IVF rates remain low. Our first aim was to verify a beneficial effect of the OEC on equine IVF. In mammals, oviductal proteins have been shown to interact with gametes and play a role in fertilization. Thus, our second aim was to identify the proteins involved in fertilization in the horse.Methods & resultsIn the first experiment, we co-incubated fresh equine spermatozoa treated with calcium ionophore and in vitro matured equine oocytes with or without porcine OEC. We showed that the presence of OEC increases the IVF rates. In the subsequent experiments, we co-incubated equine gametes with OEC and we showed that the IVF rates were not significantly different between 1) gametes co-incubated with equine vs porcine OEC, 2) intact cumulus-oocyte complexes vs denuded oocytes, 3) OEC previously stimulated with human Chorionic Gonadotropin, Luteinizing Hormone and/or oestradiol vs non stimulated OEC, 4) in vivo vs in vitro matured oocytes.In order to identify the proteins responsible for the positive effect of OEC, we first searched for the presence of the genes encoding oviductin, osteopontin and atrial natriuretic peptide A (ANP A) in the equine genome. We showed that the genes coding for osteopontin and ANP A are present. But the one for oviductin either has become a pseudogene during evolution of horse genome or has been not well annotated in horse genome sequence. We then showed that osteopontin and ANP A proteins are present in the equine oviduct using a surface plasmon resonance biosensor, and we analyzed their expression during oestrus cycle by Western blot. Finally, we co-incubated equine gametes with or without purified osteopontin or synthesized ANP A. No significant effect of osteopontin or ANP A was observed, though osteopontin slightly increased the IVF rates.ConclusionOur study shows a beneficial effect of homologous and heterologous oviduct cells on equine IVF rates, though the rates remain low. Furthers studies are necessary to identify the proteins involved. We showed that the surface plasmon resonance technique is efficient and powerful to analyze molecular interactions during fertilization.

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“…Its role is of particular importance during the process of fertilization. Oviduct epithelial cells co-culture promotes in vitro production of embryos in human (Bongso et al 1991, Kervancioglu et al 1997, bovine (Chian & Sirard 1995, Way et al 1997, Martus et al 1998, porcine (Romar et al 2001, McCauley et al 2003, deer (Locatelli et al 2005), and dromedary (Khatir et al 2004) species. Moreover, some oviductal proteins have been shown to interact with gametes and to improve efficiency of IVF in porcine (McCauley et al 2003, bovine (Martus et al 1998), human (O' Day-Bowman et al 1996), and equine (Mugnier et al 2009) species.…”

Section: Introductionmentioning

confidence: 99%

Deleted in malignant brain tumor 1 is secreted in the oviduct and involved in the mechanism of fertilization in equine and porcine species

Ambruosi¹,

Accogli²,

Douet³

et al. 2013

Reproduction

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Oviductal environment affects preparation of gametes for fertilization, fertilization itself, and subsequent embryonic development. The aim of this study was to evaluate the effect of oviductal fluid and the possible involvement of deleted in malignant brain tumor 1 (DMBT1) on IVF in porcine and equine species that represent divergent IVF models. We first performed IVF after pre-incubation of oocytes with or without oviductal fluid supplemented or not with antibodies directed against DMBT1. We showed that oviductal fluid induces an increase in the monospermic fertilization rate and that this effect is canceled by the addition of antibodies, in both porcine and equine species. Moreover, pre-incubation of oocytes with recombinant DMBT1 induces an increase in the monospermic fertilization rate in the pig, confirming an involvement of DMBT1 in the fertilization process. The presence of DMBT1 in the oviduct at different stages of the estrus cycle was shown by western blot and confirmed by immunohistochemical analysis of ampulla and isthmus regions. The presence of DMBT1 in cumulus-oocyte complexes was shown by western blot analysis, and the localization of DMBT1 in the zona pellucida and cytoplasm of equine and porcine oocytes was observed using immunofluorescence analysis and confocal microscopy. Moreover, we showed an interaction between DMBT1 and porcine spermatozoa using surface plasmon resonance studies. Finally, a bioinformatic and phylogenetic analysis allowed us to identify the DMBT1 protein as well as a DMBT1-like protein in several mammals. Our results strongly suggest an important role of DMBT1 in the process of fertilization.

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Production of dromedary (Camelus dromedarius) embryos by IVM and IVF and co-culture with oviductal or granulosa cells

Cited by 49 publications

References 24 publications

In vitro production of Sudanese camel (Camelus dromedarius) embryos from epididymal spermatozoa and follicular oocytes of slaughtered animals

In vitro production of Sudanese camel (Camelus dromedarius) embryos from epididymal spermatozoa and follicular oocytes of slaughtered animals

The secretions of oviduct epithelial cells increase the equine in vitrofertilization rate: are osteopontin, atrial natriuretic peptide A and oviductin involved?

Deleted in malignant brain tumor 1 is secreted in the oviduct and involved in the mechanism of fertilization in equine and porcine species

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