Combined treatment with cysteamine and leukemia inhibitory factor promotes guinea pig oocyte meiosis in vitro

半胱胺与白血病抑制因子联合治疗促进豚鼠卵母细胞体外减数分裂

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作者:Jiqiang Wang, Zhihui Liu, Qianru Sun, Shuang Xia, Jing Cui, Lan Yang, Liyou An, Jiaxin Zhang, Lei Su, Youqiang Su, Fuliang Du

Abstract

The guinea pig is an excellent but underused animal model due to its reproductive biology, which poses difficulties in inducing superovulation, embryo manipulation in vitro, and embryo transfer. We examined the effects of cysteamine (Cys), leukemia inhibitory factor (LIF), and Y27632 on guinea pig oocyte in vitro maturation (IVM). Cumulus-oocyte complexes were collected from antral follicles and classified into three different types before IVM. Among type I oocytes, maturation rates to metaphase II (MII) were similar in basal maturation medium and medium supplemented with Cys or LIF (39.5-40.9%), but combined Cys and LIF treatment increased the MII rate to 61.8%. Supplementation with Y27632 alone or in combination with Cys and LIF dramatically reduced the MII rate (27.7-29.7%). Similar trends were observed for type II oocytes, although their overall MII rate was lower than that of type I oocytes. The MII rate was higher among oocytes collected from 2-month-old guinea pigs compared with those from 4-month-old guinea pigs (56.5 vs. 44.8%). The optimal IVM duration was 24 h (52.5%), as 36 or 48 h of IVM reduced the MII rate (32.8-42.5%). Furthermore, Y27632 reduced the presence of microfilaments in oocytes. These findings indicate that combined supplementation of maturation medium with Cys and LIF, but not Y27632, improves the maturation efficiency of guinea pig oocytes. This study provides an important scientific basis for further efforts toward guinea pig in vitro fertilization, cloning, and gene editing by establishing an animal model for human reproduction and related diseases.

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