Proteolytically Degraded Alginate Hydrogels and Hydrophobic Microbioreactors for Porcine Oocyte Encapsulation

Gorczyca, Gabriela; Wartalski, Kamil; Tabarowski, Zbigniew; Duda, Małgorzata

doi:10.3791/61325

Cited by 6 publications

(5 citation statements)

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“…In a previous study of the same group, a similar blastocyst yield was obtained after maturation of ovine oocytes in polytetrafluoroethylene marbles compared with group culture in 2D [33]. On the other hand, in pig oocytes, although embryo development has not been assessed, Gorczyca et al (2020) [24] performed ultrastructural evaluation of COCs after IVM in fluorinated ethylene propylene marbles, and interestingly, liquid marbles were able to maintain the 3D organization by preventing their flattening and consequent disruption of gap junctions. It is noteworthy that both ovine and porcine results were obtained in serum-containing maturation medium.…”

Section: Discussionmentioning

confidence: 80%

“…Likewise, 3D systems improve cell communication and soluble factors are more stable in those systems compared with 2D systems [11,22]. In terms of in vitro embryo production, studies on 3D systems were designed to support oocyte IVM in several species using alginate microbeads [23], alginate hydrogels [24], glass scaffolds [25], agarose matrix [26], or the hanging drop method [27]. Recently, so-called liquid marbles (LM) have also become interesting as 3D-shaped bioreactors.…”

Section: Introductionmentioning

confidence: 99%

“…This system has been used for culturing microorganisms [29], embryoid bodies [30,31], or olfactory ensheathing cell spheroids [32]. The use of liquid marbles during ovine IVM showed slightly better blastocyst rates using polytetrafluoroethylene [33] and silica powder [34], while in porcine, fluorinated ethylene propylene liquid marbles were able to maintain the 3D organization of the COCs [24]. However, liquid marbles have not been validated yet in the bovine model.…”

Section: Introductionmentioning

confidence: 99%

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Alternative Culture Systems for Bovine Oocyte In Vitro Maturation: Liquid Marbles and Differentially Shaped 96-Well Plates

Fernández-Montoro

Angel‐Velez

Benedetti

et al. 2023

Animals

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In vivo-matured oocytes exhibit higher developmental competence than those matured in vitro but mimicking the in vivo environment by in vitro conditions has been challenging. Until now, conventional two-dimensional (2D) systems have been used for in vitro maturation of bovine cumulus-oocytes-complexes (COCs). However, using such systems present certain limitations. Therefore, alternative low-cost methodologies may help to optimize oocyte in vitro maturation. Here, we used two different systems to culture COCs and evaluate their potential influence on embryo development and quality. In the first system, we used treated fumed silica particles to create a 3D microenvironment (liquid marbles; LM) to mature COCs. In the second system, we cultured COCs in 96-well plates with different dimensions (flat, ultra-low attachment round-bottom, and v-shaped 96-well plates). In both systems, the nuclear maturation rate remained similar to the control in 2D, showing that most oocytes reached metaphase II. However, the subsequent blastocyst rate remained lower in the liquid marble system compared with the 96-well plates and control 2D systems. Interestingly, a lower total cell number was found in the resulting embryos from both systems (LM and 96-well plates) compared with the control. In conclusion, oocytes matured in liquid marbles or 96-well plates showed no remarkable change in terms of meiotic resumption. None of the surface geometries influenced embryo development while oocyte maturation in liquid marbles led to reduced embryo development. These findings show that different geometry during maturation did not have a large impact on oocyte and embryo development. Lower embryo production after in vitro maturation in liquid marbles was probably detected because in vitro maturation was performed in serum-free medium, which makes oocytes more sensitive to possible toxic effects from the environment.

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Section: Discussionmentioning

confidence: 80%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Alternative Culture Systems for Bovine Oocyte In Vitro Maturation: Liquid Marbles and Differentially Shaped 96-Well Plates

Fernández-Montoro

Angel‐Velez

Benedetti

et al. 2023

Animals

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“…This system has been used for culturing microorganisms [28], embryoid bodies [29,30] or olfactory ensheathing cell spheroids [31]. Liquid marbles have been also applied in ovine [32] and porcine [23] oocyte IVM but it has not been validated yet in the bovine model. Cellular responses are influenced by topographical surface features.…”

Section: Introductionmentioning

confidence: 99%

“…Likewise, 3D systems improve cell communication and soluble factors are more stable in those systems compared to 2D systems [20,21]. In terms of in vitro embryo production, studies on 3D systems were designed to support oocyte IVM in several species using alginate microbeads [22], alginate hydrogels [23], glass scaffolds [24], agarose matrix [25], or the hanging drop method [26]. Recently, LM have also become interesting as 3D bioreactors.…”

Section: Introductionmentioning

confidence: 99%

Alternative culture systems for bovine oocytein vitromaturation: liquid marbles and differentially shaped 96-well plates

Fernández-Montoro¹,

Angel‐Velez²,

Benedetti³

et al. 2022

Preprint

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In vivo matured oocytes exhibit higher developmental competence than those matured in vitro, but mimicking the in vivo environment by in vitro conditions has been challenging. Till now, conventional two-dimensional (2D) systems have been used for in vitro maturation of bovine cumulus-oocytes-complexes (COCs). However, using such systems may cause cell flattening and does not allow cumulus expansion in all dimensions, which is less physiological. Therefore, implementing a low-cost and highly effective in vivo-like microenvironment methodology may help to optimize oocyte in vitro maturation. Here, we used two different systems to culture COCs and evaluate their potential influence on embryo development and quality. In the first system, we used treated fumed silica particles to create a 3D microenvironment (liquid marbles; LM) to mature COCs. In the second system, we cultured COCs in 96-well plates with different dimensions (flat, ultra-low attachment round-bottom, and V-shaped 96-well plates). In both systems, the nuclear maturation rate remained similar to the control in 2D, showing that most oocytes reached metaphase II. However, the subsequent blastocyst rate remained lower in the liquid marble system compared to 96-well plates and control 2D systems. Interestingly, a lower total cell number was found in the resulting embryos from both systems (LM and 96-well plates) compared to the control. In conclusion, oocytes matured in liquid marbles or 96-well plates showed no remarkable change in terms of meiotic resumption, embryo development, and quality in both systems. None of the surface geometries influenced embryo development. These findings provide important inferences in many aspects of oocyte and embryo development. Further investigation is needed to determine other aspects like toxicity testing and ultrastructural changes in oocytes.

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A Simplified Method for Three-Dimensional (3D) Porcine Preantral Follicles Culture Utilizing Hydrophobic Microbioreactors

Duda

Gizler

Gorczyca

2021

Methods in Molecular Biology

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Proteolytically Degraded Alginate Hydrogels and Hydrophobic Microbioreactors for Porcine Oocyte Encapsulation

Cited by 6 publications

References 0 publications

Alternative Culture Systems for Bovine Oocyte In Vitro Maturation: Liquid Marbles and Differentially Shaped 96-Well Plates

Alternative Culture Systems for Bovine Oocyte In Vitro Maturation: Liquid Marbles and Differentially Shaped 96-Well Plates

Alternative culture systems for bovine oocytein vitromaturation: liquid marbles and differentially shaped 96-well plates

A Simplified Method for Three-Dimensional (3D) Porcine Preantral Follicles Culture Utilizing Hydrophobic Microbioreactors

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