Bebbere Daniela, Nieddu Stefano Mario, Ariu Federica, Piras Davide, Ledda Sergio
Department of Veterinary Medicine, University of Sassari, 07100 Sassari, Italy.
Cell Dynamics S.r.l., Via Viola Cielia 62/d, 40026 Imola, Italy.
Biology (Basel). 2021 Oct 25;10(11):1101. doi: 10.3390/biology10111101.
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%; = 0.066). The expression of , 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.
体外卵母细胞成熟(IVM)是一项成熟的技术。尽管在大多数哺乳动物物种中获得了较高的IVM率,但IVM卵母细胞的发育能力仍不理想。本研究的目的是评估液体大理石微生物反应器(LM)作为一种三维培养系统,对青春期前绵羊卵母细胞进行体外成熟培养的潜在有益效果,这些卵母细胞可作为内在能力较低的卵母细胞模型。青春期前绵羊卵巢的卵丘-卵母细胞复合体在含有疏水气相二氧化硅纳米颗粒的LM系统中进行体外成熟培养(LM组),或在标准条件下培养(4W对照组)。我们评估了:(a)成熟率;(b)体外受精(IVF)和胚胎培养后的发育率;(c)一组基因的表达。LM组和4W组的IVM和IVF率相似,而体外发育至囊胚阶段接近显著差异(4W组:14.1% vs. LM组:28.3%;P = 0.066)。DNA甲基化重编程相关酶以及皮质下母体复合体相关基因的表达受IVM系统影响,而在细胞应激反应方面未观察到差异。LM微生物反应器提供了一个合适的微环境来诱导青春期前绵羊卵母细胞的IVM,在包括人类在内的其他物种中,也应考虑使用该系统来提高潜力降低的卵母细胞的发育能力。