Maturation and fertilization of African lion (Panthera leo) oocytes after vitrification

Zahmel, Jennifer; Jänsch, Stefanie; Jewgenow, Katarina; Sandgreen, Ditte-Mari; Simonsen, Kim Skalborg; Colombo, Martina

doi:10.1016/j.cryobiol.2020.11.011

Cited by 14 publications

(12 citation statements)

References 55 publications

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“…These animals, aged 5 days, 10 weeks (2 animals), and 7 and 17 years, were therefore not included in this survey. Immediately after isolation, 874 lion oocytes were subjected to culture, whereas 59 good quality oocytes were frozen by vitrification (see specific results in [ 19 ]). After 24–36 h of IVM, 355 oocytes or 40.6% were classified as mature either by determining the nuclear stage with PI or the visibility of extruded polar bodies or by cleavage after fertilization.…”

Section: Resultsmentioning

confidence: 99%

“…Oocytes and embryos were vitrified by the Cryotop method as previously described [ 19 ]. Briefly, one to eight oocytes were equilibrated at room temperature in an equilibration solution (ES) containing 7.5% ( v / v ) ethylene glycol (EG) and 7.5% dimethyl sulfoxide (Me 2 SO) in Medium 199 with 20% fetal bovine serum (FBS) for 15 min.…”

Section: Methodsmentioning

confidence: 99%

“…Laparoscopic oocyte retrieval and subsequent fertilization has been performed [ 17 ], and as part of the Felid-Gamete-Rescue-Project our research group was able to show that also in vitro maturation followed by intracytoplasmic sperm injection led to successful fertilization and embryonic development up to the blastocyst stage [ 18 ]. We were also able to cryopreserve lion oocytes for the first time [ 19 ]. The oocytes survived vitrification, matured in vitro, and cleaved subsequent to in vitro fertilization.…”

Section: Introductionmentioning

confidence: 99%

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Current State of In Vitro Embryo Production in African Lion (Panthera leo)

Zahmel

Simonsen

Stagegaard³

et al. 2022

Animals

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In the last 30–40 years, in vitro maturation (IVM) and fertilization (IVF) of domestic cat oocytes have been established as part of the panel of assisted reproduction technologies. As a representative of wild felids, the African lion is not yet considered endangered. Nevertheless, the zoo population management of the African lion itself as well as other closely related felids would benefit from the establishment of an IVF system. Here, we aimed to investigate the transferability of domestic cat IVF technology to the African lion. From the ovaries of 42 lionesses aged between 0.75 and 15 years, a total of 933 IVF-suitable oocytes were retrieved and subjected to IVM and IVF. The overall maturation rate was 40.6% and 18.9% of these oocytes cleaved after fertilization, respectively. Embryos were generated by intracytoplasmic sperm cell injection as well as co-culture with epididymal sperm. Improvements in the model system also led to an improved outcome with in vitro produced embryos in the lion. Compared to domestic cats, the transportation of gonads to a specialized laboratory was time-consuming and influenced oocyte quality negatively. In conclusion, the domestic cat IVF system is adoptable for the African lion, although success rates are still lower.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Current State of In Vitro Embryo Production in African Lion (Panthera leo)

Zahmel

Simonsen

Stagegaard³

et al. 2022

Animals

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“…In recent years, cryopreservation of oocytes has been widely applied in preservation of animal germplasm, developmental biology research, and assisted reproduction among others (1)(2)(3). In animal production, this approach could be used to establish oocyte banks, facilitating animal production and breeding programs (4), and also preserving diversity of livestock genetic resources (5)(6)(7)(8).…”

Section: Introductionmentioning

confidence: 99%

“…(1) Expression level of TOMM20 and MMP were detected in MII oocytes by immunofluorescence staining, and ATP content was measured using a multi-plate reader containing chemiluminescence in MII oocytes. (2) Levels of mRNA of mitochondrial biogenesis related genes (Sirt1, Pgc-1α, Tfam) were detected in MII oocytes with Q-PCR.Experiment 3: Effects of melatonin on oxidative stress and apoptosis in vitrified-warmed mouse GV oocytes through regulation of p-Drp1 Ser616 (1). The ROS level, early apoptosis and Cytochrome C content were detected in MII oocytes by immunofluorescence staining.…”

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confidence: 99%

Melatonin Promotes in vitro Development of Vitrified-Warmed Mouse GV Oocytes, Potentially by Modulating Phosphorylation of Drp1

et al. 2021

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Previous studies have shown that melatonin can mitigate cryopreservation-induced mitochondrial dysfunction in oocytes; however, the underlying molecular mechanism remains unclear. The objective of the present study was to investigate whether melatonin can improve the mitochondrial function during in vitro maturation of vitrified-warmed mouse germinal vesicle (GV) oocytes by modulating phosphorylation of dynamin related protein 1 (Drp1). Vitrification/warming procedures resulted in the following: (1) After cryopreservation of mouse GV oocytes, the phosphorylation level of Drp1 at Ser616 (p-Drp1 Ser616) in metaphase II (MII) oocytes was increased (P < 0.05). Furthermore, the rates of in vitro maturation, cleavage and blastocyst formation after parthenogenetic activation were decreased (P < 0.05). (2) In MII oocytes, the expression levels of translocase of the mitochondrial outer membrane 20 (TOMM20), mitochondrial membrane potential (MMP), adenosine triphosphate (ATP) content, and mRNA levels of mitochondrial biogenesis-related genes (Sirt1, Pgc-1α, Tfam) were all decreased (P < 0.05), and (3) Reactive oxygen species (ROS) level, early apoptosis level, Cytochrome C release and mRNA levels of pro-apoptotic related genes (Bax, Caspase9, Caspase3) in MII oocytes were all increased (P < 0.05). The results of this study further revealed that negative impacts of GV oocyte cryopreservation were mitigated by supplementation of warming and in vitro maturation media with 10−7mol /L melatonin or 2 x 10−5mol/L Mdivi-1 (Drp1 inhibitor). Therefore, we concluded that 10−7mol/L melatonin improved mitochondrial function, reduced oxidative stress and inhibited apoptosis by regulating phosphorylation of Drp1, thereby enhancing in vitro development of vitrified-warmed mouse GV oocytes.

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Gross¹

2021

Nachr Chem

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Maturation and fertilization of African lion (Panthera leo) oocytes after vitrification

Cited by 14 publications

References 55 publications

Current State of In Vitro Embryo Production in African Lion (Panthera leo)

Current State of In Vitro Embryo Production in African Lion (Panthera leo)

Melatonin Promotes in vitro Development of Vitrified-Warmed Mouse GV Oocytes, Potentially by Modulating Phosphorylation of Drp1

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