The variable success of in vitro maturation: can we do better?

Luciano, A.M.; Franciosi, Federica; Barros, Rodrigo Garcia; Dieci, Cecilia; Lodde, Valentina

doi:10.21451/1984-3143-ar2018-0021

Cited by 16 publications

(10 citation statements)

References 100 publications

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“…Furthermore, cellular integrity, determined by cytoskeleton analysis and cell organization, is suggested to be an indicator of embryo quality in other species [20,21]. It is evident that there is a need to more clearly define the contribution of these parameters to embryo quality, specifically during in vitro production where embryo viability appears to be compromised [22][23][24][25].…”

Section: Introductionmentioning

confidence: 99%

Improved cryopreservation of in vitro produced bovine embryos using FGF2, LIF, and IGF1

et al. 2021

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In vitro embryo production systems are limited by their inability to consistently produce embryos with the competency to develop to the blastocyst stage, survive cryopreservation, and establish a pregnancy. Previous work identified a combination of three cytokines [fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF), and insulin-like growth factor 1 (IGF1)], called FLI, that we hypothesize improve preimplantation development of bovine embryos in vitro. To test this hypothesis, FLI was supplemented into oocyte maturation or embryo culture medium. Embryos were produced in vitro using abattoir-derived oocytes and fertilized with sperm from a single bull known to have high fertility. After an 18–20 h fertilization period, putative zygotes were cultured in synthetic oviductal fluid (SOF) for 8 days. The addition of FLI to the oocyte maturation medium increased (P < 0.05) the dissociation of transzonal projections at 12, 18, and 24 h of maturation, as well as, the proportion of oocytes that reached the metaphase II stage of meiosis. Additionally, lipid content was decreased (P < 0.05) in the blastocyst stage embryo. The addition of FLI during the culture period increased development to the blastocyst stage, cytoskeleton integrity, and survival following slow freezing, as well as, decreased post thaw cell apoptosis (P < 0.05). In conclusion, the supplementation of these cytokines in vitro has the potential to alleviate some of the challenges associated with the cryo-survival of in vitro produced bovine embryos through improving embryo development and embryo quality.

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Section: Introductionmentioning

confidence: 99%

Improved cryopreservation of in vitro produced bovine embryos using FGF2, LIF, and IGF1

et al. 2021

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“…Although perspectives are promising, the clinical outcomes of IVM oocytes are lower compared with those of oocytes matured in vivo [ 17 , 18 , 19 , 20 , 21 ]. To improve IVM protocols, in vitro culture conditions should be optimized to better support nuclear and cytoplasmic modifications occurring physiologically as a result of in vivo ovulatory stimuli [ 22 , 23 , 24 , 25 , 26 ].…”

Section: Introductionmentioning

confidence: 99%

Investigating and Modelling an Engineered Millifluidic In Vitro Oocyte Maturation System Reproducing the Physiological Ovary Environment in the Sheep Model

Mastrorocco

Cacopardo²,

Temerario³

et al. 2022

Cells

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In conventional assisted reproductive technologies (ARTs), oocytes are in vitro cultured in static conditions. Instead, dynamic systems could better mimic the physiological in vivo environment. In this study, a millifluidic in vitro oocyte maturation (mIVM) system, in a transparent bioreactor integrated with 3D printed supports, was investigated and modeled thanks to computational fluid dynamic (CFD) and oxygen convection-reaction-diffusion (CRD) models. Cumulus-oocyte complexes (COCs) from slaughtered lambs were cultured for 24 h under static (controls) or dynamic IVM in absence (native) or presence of 3D-printed devices with different shapes and assembly modes, with/without alginate filling. Nuclear chromatin configuration, mitochondria distribution patterns, and activity of in vitro matured oocytes were assessed. The native dynamic mIVM significantly reduced the maturation rate compared to the static group (p < 0.001) and metaphase II (MII) oocytes showed impaired mitochondria distribution (p < 0.05) and activity (p < 0.001). When COCs were included in a combination of concave+ring support, particularly with alginate filling, oocyte maturation and mitochondria pattern were preserved, and bioenergetic/oxidative status was improved (p < 0.05) compared to controls. Results were supported by computational models demonstrating that, in mIVM in biocompatible inserts, COCs were protected from shear stresses while ensuring physiological oxygen diffusion replicating the one occurring in vivo from capillaries.

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“…However, the overall efficiency of the in vitro production process remains low (Lonergan & Fair, 2016; Luciano et al, 2018; Marsico et al, 2019). Additionally, the lack of homogeneity of postcryopreservation survival results of IVP embryos continues to hinder the massive dissemination of this technology.…”

Section: Introductionmentioning

confidence: 99%

Transcriptional profiling of embryo cryotolerance

Marsico

Caetano

Rodrigues³

et al. 2020

Molecular Reproduction Devel

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The cryosurvival of embryos is a complex process involving dynamic and integrated morphological, functional, and molecular changes. Here, we evaluated the transcriptional profiling of bovine embryos possessing high and low cryotolerance (HC and LC, respectively) by assessing the resumption of development. Embryos were produced in vitro (N = 1137) and cryopreserved (N = 894). Blastocysts samples possessed pronounced group individualization at RNA sequencing. A total of 114 genes were differentially expressed, and 27 and 84 genes were upregulated in HC and LC, respectively. Among the over-represented biological functions, cellular growth and proliferation, cell death and survival, and organismal survival were predicted to be activated, while cellular movement and cell-to-cell signaling were predicted to be inhibited in HC embryos. Enriched canonical pathways and upstream regulators related to cellular proliferation and survival (HC), inflammatory processes, and cell death (LC) were predicted to represent two embryonic molecular profiles present during the resumption of development after cryopreservation. The marked contrast in transcriptional profiles between HC and LC strongly suggests the influence of embryonic competence after cryopreservation on its respective transcriptome and indicated that HC and LC presented two different molecular strategies to overcome cryopreservation-related stress and resume postcryopreservation development.

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The variable success of in vitro maturation: can we do better?

Cited by 16 publications

References 100 publications

Improved cryopreservation of in vitro produced bovine embryos using FGF2, LIF, and IGF1

Improved cryopreservation of in vitro produced bovine embryos using FGF2, LIF, and IGF1

Investigating and Modelling an Engineered Millifluidic In Vitro Oocyte Maturation System Reproducing the Physiological Ovary Environment in the Sheep Model

Transcriptional profiling of embryo cryotolerance

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