Abstract
The in vitro maturation of oocytes is frequently used as an assisted reproductive technique (ART), and has been successfully established in humans and rodents. To overcome the limitations of ART, novel procedures for the in vitro maturation of early follicles are emerging. During the follicle isolation procedure, the unintended rupture of each follicle leads to a release of extra oocytes. Such oocytes, which are obtained during follicle isolation from marmosets, can be used for early maturation studies. Marmoset (Callithrix jacchus), which is classified as a new-world monkey, is a novel model that has been employed in reproductive biomedical research, as its reproductive physiology is similar to that of humans in several aspects. The ovaries of female marmosets were collected, and the excess oocytes present during follicle isolation were retrieved without pre-gonadotropin induction. Each oocyte was matured in vitro for 48 h in the presence of various concentrations of human chorionic gonadotropin (hCG) and epidermal growth factor (EGF), and the maturity of oocytes and optimal maturation conditions were evaluated. Each oocyte was individually reverse-transcribed, and the expression of mRNAs and microRNAs (miRs) were analyzed. Concentrations of hCG significantly affected the maturation rate of oocytes [the number of metaphase II (MII) oocytes]. The expression of BMP15 and ZP1 was highest when the oocytes were matured using 100 IU/L of hCG without pre-treatment with gonadotropins, and that of Cja-mir-27a was highest when cultured with follicle stimulating hormone. These results suggest that these up-regulated miRs affect the maturation of oocytes. Interactions with other protein networks were analyzed, and a strong association of BMP15 and ZP1 with sperm binding receptor (ACR), anti-Müllerian hormone (AMH), and AMH receptor was demonstrated, which is related to the proliferation of granulosa cells. Collectively, on the basis of these results, the authors propose optimal maturation conditions of excess oocytes of marmoset without in vivo gonadotropin treatment, and demonstrated the roles of miRs in early oocyte maturation at the single-cell level in marmosets.
Highlights
Oocyte in vitro maturation (IVM) is a popular technology in assisted reproductive technology (ART)
Numerous studies focused on the maturation conditions and factors regulating or enhancing developmental competence for a higher yield of in vitro maturation and transition to early stage embryos, e.g., vitamin D [25,26] and inositol [27,28]
Non-human primates can be classified into old-world and new-world monkeys [29]; the marmoset belongs to the class of new-world monkeys
Summary
Oocyte in vitro maturation (IVM) is a popular technology in assisted reproductive technology (ART). Environmental and otherwise, affecting the developmental competency of oocytes have been studied using many species, such as mice and domestic animals; such animals have different reproductive characteristics than those of humans [2,3,4,5,6,7]. These animals have estrous cycles and multi-fetal pregnancies, while humans undergo a menstrual cycle and mostly have a single-fetal pregnancy. During the isolation of follicles for IVFM, the use of oocytes from the unintended rupture of early follicles could be another strategy that could be studied and developed using marmoset as a reproductive study model
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