The never‐ending search of an ideal culture system for domestic cat oocytes and embryos

Luvoni, G.C.; Colombo, Martina; Morselli, Maria Giorgia

doi:10.1111/rda.13331

Cited by 10 publications

(10 citation statements)

References 97 publications

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“…Since the extracellular environment might positively influence the intracellular conditions [21], the culture of VOs in a 3D microenvironment, more similar to the follicular niche, was tested. Although the in vitro development of immature VOs after warming is still a challenge, in this study VOs were able to mature and be fertilized, and similar maturation and embryonic developmental rates were obtained in both 3D and 2D conditions ( p > 0.05), pointing out that 3D barium alginate microcapsules, closer to physiological conditions, are suitable for the culture of VOs, although not enough for these highly demanding gametes.…”

Section: Discussionmentioning

confidence: 99%

“…To ameliorate oocytes performances in vitro, other successful attempts have been made using coculture systems, so that the presence of somatic cell monolayers or competent cumulus-oocyte complexes (COCs) gave physical (structural support) or chemical (secreted paracrine factors) improvements, which are beneficial for cocultured oocytes [18,19,20,21]. The association of 3D systems and cocultures was applied with good results for fresh domestic cat oocytes [22,23], but for VOs, satisfactory improvements in terms of maturation and cleavage rates have only been documented in 2D coculture in other species (pig [24,25], buffalo [26]).…”

Section: Introductionmentioning

confidence: 99%

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Developmental Competence of Domestic Cat Vitrified Oocytes in 3D Enriched Culture Conditions

Colombo

Morselli

Tavares

et al. 2019

Animals

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Cryoinjuries severely affect the competence of vitrified oocytes (VOs) to develop into embryos after warming. The use of culture conditions that provide physical and chemical support and resemble the in vivo microenvironment in which oocytes develop, such as 3D scaffolds and coculture systems, might be useful to improve VOs outcomes. In this study, an enriched culture system of 3D barium alginate microcapsules was employed for the in vitro embryo production of domestic cat VOs. Cryotop vitrified-warmed oocytes were in vitro matured for 24 h in the 3D system with or without fresh cumulus-oocyte complexes (COCs) in coculture, whereas a control group of VOs was cultured in traditional 2D microdrops of medium. After in vitro fertilization, presumptive embryos were cultured in 3D or 2D systems according to the maturation conditions. Vitrified oocytes were able to mature and develop into embryos in 3D microcapsules (17.42 ± 11.83%) as well as in 2D microdrops (14.96 ± 8.80%), but the coculture with companion COCs in 3D resulted in similar proportions of VOs embryo development (18.39 ± 16.67%; p = 1.00), although COCs presence allowed for blastocyst formation (0.95 ± 2.52%). In conclusion, embryos until late developmental stages were obtained from cat VOs, and 3D microcapsules were comparable to 2D microdrops, but improvements in post-warming conditions are still needed.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Developmental Competence of Domestic Cat Vitrified Oocytes in 3D Enriched Culture Conditions

Colombo

Morselli

Tavares

et al. 2019

Animals

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“…Oocytes were singly in vitro matured for 24 h in a controlled atmosphere (39°C and 5% CO 2 in air) in 20 μL microdrops of IVM medium ( 33 , 34 ) [Medium 199 supplemented with 3 mg/mL BSA, 0.1 mg/mL cysteine, 1.4 mg/mL HEPES, 0.25 mg/mL sodium pyruvate, 0.6 mg/mL sodium lactate, 0.15 mg/mL L-glutamine, 0.055 mg/mL gentamicin, 0.2 IU/mL human luteinizing hormone (LH), and 0.5 IU/mL human pituitary follicle-stimulating hormone (hFSH)] covered by mineral oil in Petri dishes.…”

Section: Methodsmentioning

confidence: 99%

“…Based on the results of Experiment I, warmed oocytes were incubated for 24 h before staining. Oocytes were singly in vitro matured for 24 h in a controlled atmosphere (39 • C and 5% CO 2 in air) in 20 µL microdrops of IVM medium (33,34) [Medium 199 supplemented with 3 mg/mL BSA, 0.1 mg/mL cysteine, 1.4 mg/mL HEPES, 0.25 Images were captured in black and white and pseudo-colored after acquisition with the Imaging Software ZEN 2.5 blue edition. Black and white balance of Hoechst (1) and TUNEL (2) images was adjusted after coloring to make nuclear stainings more visible in print.…”

Section: Staining For Apoptotic Signal Detection (Experiments I and Ii)mentioning

confidence: 99%

Inhibition of Apoptotic Pathways Improves DNA Integrity but Not Developmental Competence of Domestic Cat Immature Vitrified Oocytes

et al. 2020

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Being a model for endangered wild felids, cryopreservation protocols for domestic cat oocytes are under continuous development. Immature vitrified oocytes (VOs) are a valuable resource for fertility preservation programs, but they often degenerate after warming and their in vitro development is poor. Since the exact mechanisms are not clear, this study assessed whether vitrification might trigger two apoptotic markers (DNA fragmentation and caspase activity, Experiment I) and the effects of a chemical inhibitor (i.e., the pan-caspase inhibitor Z-VAD-FMK) on the same markers (Experiment II) and on VOs in vitro development (Experiment III). The overarching aim was to check whether apoptosis inhibition might be a strategy to improve cat oocytes cryotolerance. In Experiment I, vitrification induced DNA fragmentation and increased caspase activity in VOs incubated for 24 h after warming (DNA fragmentation: 59.38%; caspase activity: 414.6 ± 326.8) compared to a fresh control (9.68%; 199.6 ± 178.3; p = 0.02). In Experiment II, the addition of Z-VAD-FMK to vitrification-warming and incubation media decreased DNA fragmentation and caspase activity (8.82%; 243.7 ± 106.9) compared to control (untreated) VOs (69.44%; 434.5 ± 248.3; p < 0.001). In Experiment III, Z-VAD-FMK brought maturation rates of treated VOs close to those of fresh oocytes (53.13 and 65.38%, respectively, p = 0.057), but there were no differences in VOs embryo development (cleavage rates; Z-VAD-FMK-treated VOs: 34.38%; control VOs: 31.78%; p = 0.69). In summary, vitrification increased apoptotic markers in cat VOs, and while Z-VAD-FMK was able to hinder DNA damage and caspase activity, its addition was not determinant for embryo development. To make the best use of VOs, other oocyte in vitro maturation and embryo culture strategies, such as the addition of other inhibitors or their prolonged use, should be investigated.

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“…Since the beginning of feline assisted reproductive techniques (ART) in the 1970s (Hamner, Jennings, & Sojka, ), much work has been done to develop in vitro fertilization techniques in cats, including the optimization of culture conditions (reviewed by Luvoni, Colombo, & Morselli, ). Everything that an embryo needs for its growth and development—a stable environment and nutrients—is provided by the medium in which it is cultured.…”

Section: Introductionmentioning

confidence: 99%

The use of human and bovine commercial media for oocyte maturation and embryo development in the domestic cat (Felis catus)

Prochowska

Niżański

Partyka

et al. 2019

Reprod Domestic Animals

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The aim of this study was to examine the suitability of commercial media designed for humans and cattle for oocyte maturation and embryo culture in the domestic cat. In Exp. I, feline oocytes collected ex vivo were subjected to in vitro maturation in a laboratory-made culture medium (based on M199) or a commercial medium designed for cattle cells (BO-IVM ® ). In Exp. II, ICSI-derived feline embryos were cultured for 7 days in a commercial human (Continuous Single Culture ® ) or bovine (BO-EC ® ) cell medium. The rates of cleavage, morula and blastocyst formation were evaluated at 24 hr, 6 days and 7 days after ICSI, respectively, and compared between experimental groups. At the end of culture, embryos were assessed for viability and apoptotic changes. In Exp. I, no statistically significant difference in oocyte maturation outcome between laboratory-made (52.7%) and commercial media (58.9%) was observed. However, the use of a commercial medium prepared for use with bovine cells resulted in a significantly lower variance of the maturation rate. In Exp. II, no statistically significant differences between two commercial media were observed for cleavage (67.5% and 64.5%), morula (39.3% and 47.1%) and blastocyst rates (25.0% and 19.6%), as well as for the percentage of late apoptotic blastomeres. Morulae cultured in medium marketed for humans exhibited significantly more early apoptotic (43.2 ± 31.2% vs. 23.4 ± 23.2%) and necrotic (60.6 ± 47.6% vs. 29.4 ± 22.6%) blastomeres. In conclusion, both commercial media tested are suitable for in vitro oocyte maturation and embryo culture procedures in cats. It is remarkable that a culture medium designed for use in cattle for in vitro maturation of cat oocytes provides more reproducible results. K E Y W O R D Scat, commercial media, embryo culture, oocyte maturation

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The never‐ending search of an ideal culture system for domestic cat oocytes and embryos

Cited by 10 publications

References 97 publications

Developmental Competence of Domestic Cat Vitrified Oocytes in 3D Enriched Culture Conditions

Developmental Competence of Domestic Cat Vitrified Oocytes in 3D Enriched Culture Conditions

Inhibition of Apoptotic Pathways Improves DNA Integrity but Not Developmental Competence of Domestic Cat Immature Vitrified Oocytes

The use of human and bovine commercial media for oocyte maturation and embryo development in the domestic cat (Felis catus)

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