In vitro and in vivo development of domestic cat embryos generated by in vitro fertilization after eCG priming and oocyte in vitro maturation

Veraguas, D.; Saez, S.; Aguilera, C.; Echeverry, Diana; Gallegos, PF; Saez-Ruiz, D.; Castro, Fidel Ovídio; Rodríguez-Álvarez, L.

doi:10.1016/j.theriogenology.2020.02.012

Cited by 9 publications

(4 citation statements)

References 37 publications

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“…Testes were transported in sterile 0.9% NaCl solution with 0.1% gentamycin at room temperature. The caudal portions of the epididymis were cut into small pieces of ~1 mm in a 100 mm petri dish containing 10 mL of He199 supplemented with 0.3% fraction V BSA instead of FBS (He199-BSA) at 38.5 • C. Epididymal spermatozoa were processed and stored at 4 • C, as we previously described [150].…”

Section: Sperm Collection and In Vitro Fertilizationmentioning

confidence: 99%

“…IVF was conducted in four-well dishes with 500 µL of TALP medium supplemented with 6 mg/mL BSA, 0.36 mM sodium pyruvate, 1 mM glutamine, 2.2 mM calcium lactate, 1% MEM nonessential amino acids (NEAA), 0.5% MEM essential amino acids (EAA), 0.01 mg/mL heparin sodium salt and 50 µg/mL gentamycin (TALP-IVF). Refrigerated spermatozoa were allowed to swim up for 30 min in He199-BSA at 38.5 • C [150]. The pellet was collected and resuspended in TALP-IVF.…”

Section: Sperm Collection and In Vitro Fertilizationmentioning

confidence: 99%

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Proteomic Analysis of Domestic Cat Blastocysts and Their Secretome Produced in an In Vitro Culture System without the Presence of the Zona Pellucida

Veraguas-Dávila,

Zapata-Rojas,

Aguilera

et al. 2024

IJMS

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Domestic cat blastocysts cultured without the zona pellucida exhibit reduced implantation capacity. However, the protein expression profile has not been evaluated in these embryos. The objective of this study was to evaluate the protein expression profile of domestic cat blastocysts cultured without the zona pellucida. Two experimental groups were generated: (1) domestic cat embryos generated by IVF and cultured in vitro (zona intact, (ZI)) and (2) domestic cat embryos cultured in vitro without the zona pellucida (zona-free (ZF group)). The cleavage, morula, and blastocyst rates were estimated at days 2, 5 and 7, respectively. Day 7 blastocysts and their culture media were subjected to liquid chromatography–tandem mass spectrometry (LC–MS/MS). The UniProt Felis catus database was used to identify the standard proteome. No significant differences were found in the cleavage, morula, or blastocyst rates between the ZI and ZF groups (p > 0.05). Proteomic analysis revealed 22 upregulated and 20 downregulated proteins in the ZF blastocysts. Furthermore, 14 proteins involved in embryo development and implantation were present exclusively in the culture medium of the ZI blastocysts. In conclusion, embryo culture without the zona pellucida did not affect in vitro development, but altered the protein expression profile and release of domestic cat blastocysts.

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Section: Sperm Collection and In Vitro Fertilizationmentioning

confidence: 99%

Section: Sperm Collection and In Vitro Fertilizationmentioning

confidence: 99%

Proteomic Analysis of Domestic Cat Blastocysts and Their Secretome Produced in an In Vitro Culture System without the Presence of the Zona Pellucida

Veraguas-Dávila,

Zapata-Rojas,

Aguilera

et al. 2024

IJMS

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“…In horses, successful IVF has been achieved with a sperm concentration of 1 × 10 6 spermatozoa/mL [ 125 ], while for IVF in cattle, the sperm concentration ranges from 0.5 × 10 6 to 2 × 10 6 spermatozoa/mL [ 126 ]. In domestic cats, a sperm concentration of 1.5 × 10 6 to 2.5 × 10 6 spermatozoa/mL is typically utilized [ 127 ]. ICSI offers the advantage of direct injection of a single spermatozoon into the cytoplasm of a mature oocyte using micromanipulation, thereby eliminating the requirement for high-quality and high-quantity sperm [ 128 , 129 ].…”

Section: Artificial Reproduction Techniques (Arts)mentioning

confidence: 99%

Postmortem Collection of Gametes for the Conservation of Endangered Mammals: A Review of the Current State-of-the-Art

Huijsmans

Hassan

Smits

et al. 2023

Animals

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The collection of gametes from recently deceased domestic and wildlife mammals has been well documented in the literature. Through the utilization of gametes recovered postmortem, scientists have successfully produced embryos in 10 different wildlife species, while in 2 of those, offspring have also been born. Thus, the collection of gametes from recently deceased animals represents a valuable opportunity to increase genetic resource banks, obviating the requirement for invasive procedures. Despite the development of several protocols for gamete collection, the refinement of these techniques and the establishment of species–specific protocols are still required, taking into account both the limitations and the opportunities. In the case of wildlife, the optimization of such protocols is impeded by the scarcity of available animals, many of which have a high genetic value that must be protected rather than utilized for research purposes. Therefore, optimizing protocols for wildlife species by using domestic species as a model is crucial. In this review, we focused on the current advancements in the collection, preservation, and utilization of gametes, postmortem, in selected species belonging to Equidae, Bovidae, and Felidae, both domestic and wildlife.

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“…After IVM, the oocytes were washed twice in an IVF medium composed of TALP supplemented with heparin (10 µg/mL) (Veraguas et al, 2020). Oocytes (10-15 per drop) were co-incubated with 2 × 10 6 sperm/mL in 50 µL drops of IVF medium for 18 h at 38.5 °C and 5% CO 2 .…”

Section: Heterologous In Vitro Fertilization and Embryonic Developmentmentioning

confidence: 99%

Heterologous in vitro fertilization and embryo production for assessment of jaguar (Panthera onca Linnaeus, 1758) frozen-thawed semen in different extenders

et al. 2022

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Heterologous in vitro fertilization (IVF) is an important tool for assessing fertility of endangered mammals such as the jaguar, considering difficult access to females for artificial insemination and to obtain homologous oocytes. We aimed to evaluate the fertility of jaguar sperm cryopreserved with different extenders, using domestic cat oocytes to assess the development of hybrid embryos. Semen from four captive jaguars was obtained by electroejaculation. Samples were cryopreserved in powdered coconut water (ACP-117c) or Tris extender containing 20% egg yolk and 6% glycerol. Thawed spermatozoa were resuspended (2.0 × 10 6 spermatozoa/mL) in IVF medium and co-incubated with cat oocytes matured in vitro for 18 h. Presumptive zygotes were cultured for 7 days. After 48 h, cleavage rate was evaluated, and non-cleaved structures were stained for IVF evaluation. On days 5 and 7, the rate of morula and blastocyst formation was assessed. Data were analyzed using the Fisher exact test (p < 0.05). No difference was observed between ACP-117c and Tris extenders, respectively, for oocytes with 2nd polar body (2/51, 3.9 ± 2.9% vs. 2/56, 3.6 ± 3.1%), pronuclear structures (5/51, 9.8 ± 4.7% vs. 8/56, 14.3 ± 8.0%), and total IVF rates (7/36, 19.4 ± 5.0% vs. 10/37, 27.0 ± 13.8%). All the samples fertilized the oocytes, with 22.9 ± 3.2% (16/70) and 16.7 ± 3.6% (12/72) cleavage of mature oocytes for ACP-117c and Tris extenders, respectively. Morula rates of 4.3 ± 2.3% (3/70) and 5.6 ± 2.2% (4/72) were observed for ACP-117c and Tris, respectively. Only the Tris extender demonstrated blastocyst production (2/12, 16.7 ± 1.5% blastocyst/cleavage). We demonstrated that jaguar ejaculates cryopreserved using ACP-117c and Tris were suitable for IVF techniques, with blastocyst production by ejaculates cryopreserved in Tris. This is a first report of embryos produced in vitro using jaguar sperm and domestic cat oocytes through IVF.

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In vitro and in vivo development of domestic cat embryos generated by in vitro fertilization after eCG priming and oocyte in vitro maturation

Cited by 9 publications

References 37 publications

Proteomic Analysis of Domestic Cat Blastocysts and Their Secretome Produced in an In Vitro Culture System without the Presence of the Zona Pellucida

Proteomic Analysis of Domestic Cat Blastocysts and Their Secretome Produced in an In Vitro Culture System without the Presence of the Zona Pellucida

Postmortem Collection of Gametes for the Conservation of Endangered Mammals: A Review of the Current State-of-the-Art

Heterologous in vitro fertilization and embryo production for assessment of jaguar (Panthera onca Linnaeus, 1758) frozen-thawed semen in different extenders

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