3D Liquid Marble Microbioreactors Support In Vitro Maturation of Prepubertal Ovine Oocytes and Affect Expression of Oocyte-Specific Factors

Abstract

In vitro oocyte maturation (IVM) is a well-established technique. Despite the high IVM rates obtained in most mammalian species, the developmental competence of IVM oocytes is suboptimal. The aim of this work was to evaluate the potential beneficial effects of a liquid marble microbioreactor (LM) as a 3D culture system to mature in vitro prepubertal ovine oocytes, as models of oocytes with intrinsic low competence. Cumulus–oocyte complexes of prepubertal sheep ovaries were in vitro matured in a LM system with hydrophobic fumed-silica-nanoparticles (LM group) or in standard conditions (4W control group). We evaluated: (a) maturation and (b) developmental rates following in vitro fertilization (IVF) and embryo culture; (c) expression of a panel of genes. LM and 4W groups showed similar IVM and IVF rates, while in vitro development to blastocyst stage approached significance (4W: 14.1% vs. LM: 28.3%; p = 0.066). The expression of GDF9, of enzymes involved in DNA methylation reprogramming and of the subcortical maternal complex was affected by the IVM system, while no difference was observed in terms of cell-stress-response. LM microbioreactors provide a suitable microenvironment to induce prepubertal sheep oocyte IVM and should be considered to enhance the developmental competence of oocytes with reduced potential also in other species, including humans.

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

“…However, the toxicity of treated fumed silica particles has not been tested in oocytes. Therefore, we hypothesize that treated fumed silica particles could exhibit some toxicity in bovine oocytes since our findings differ from the results obtained by Bebbere et al, who matured ovine oocytes in LM formed with the same particles, showing better blastocyst rate compared to those matured in 2D conditions [41]. Additionally, in a previous study of the same group, better blastocyst yield was reached after the maturation of ovine oocytes in polytetrafluoroethylene marbles compared to the group in 2D [32].…”

“…We proved that meiotic resumption is not affected in bovine oocytes by the use of LM, since the proportion of oocytes that reached metaphase II was similar in both 2D- and 3D-systems, which concord with previous studies in the cat [41] and sheep [32,41]. In our study, although oocytes matured in LM were able to reach the blastocyst stage, the blastocyst yield was reduced compared to oocytes matured in 2D.…”

“…We found that both matrix and non-matrix 3D culture systems had a similar effect as 2D culture systems in terms of oocyte nuclear maturation, while embryo development was similar after oocyte maturation in the 96-well plates but lower in LM. We proved that meiotic resumption is not affected in bovine oocytes by the use of LM, since the proportion of oocytes that reached metaphase II was similar in both 2D-and 3D-systems, which concord with previous studies in the cat [41] and sheep [32,41]. In our study, although oocytes matured in LM were able to reach the blastocyst stage, the blastocyst yield was reduced compared to oocytes matured in 2D.…”

Effects of IGF-1 on the Three-Dimensional Culture of Ovarian Preantral Follicles and Superovulation Rates in Mice

“…The results of the current research have proven that designing modern and powerful approaches to the murine ex vivo 3D model of IGF-1-assisted growth and maturation of ovarian secondary preantral follicles in order to obtain satisfactory outcomes of IVM might be reliable and feasible for the purposes of achieving high efficiency rates in such novel ARTs as IVF/ICSI in humans and other mammalian species [ 65 , 66 , 67 , 68 ] or somatic cell cloning in other mammalian species [ 69 , 70 , 71 , 72 ].…”

IVM Advances for Early Antral Follicle-Enclosed Oocytes Coupling Reproductive Tissue Engineering to Inductive Influences of Human Chorionic Gonadotropin and Ovarian Surface Epithelium Coculture

“…On the other hand, the strategies tested to improve the performance of IVM of low competent oocytes were mainly focused on the use of dynamic cell culture devices (3D cultures) [ 51 , 52 ] and/or the supplementation of the IVM system with factors and/or molecules essential to the ovarian microenvironment [ 53 , 54 ]. Alternatively, the “simulated physiological oocyte maturation” (SPOM) protocol has also been applied to improve low competent oocyte IVM in bovine [ 55 ] and caprine [ 56 , 57 , 58 ] species.…”