Tonai Shingo, Nakanishi Tomoya, Yamaoka Manami, Okamoto Asako, Shimada Masayuki, Yamashita Yasuhisa
Graduate School of Comprehensive Scientific Research Prefectural University of Hiroshima Shobara Japan.
Graduate School of Integrated Sciences for Life Hiroshima University Higashi-Hiroshima Japan.
Reprod Med Biol. 2023 Aug 4;22(1):e12529. doi: 10.1002/rmb2.12529. eCollection 2023 Jan-Dec.
Since the developmental competence of oocytes cultured after in vitro maturation (IVM) is low, it is necessary to improve the IVM method for efficient offspring production. In this study, we revealed that transferrin (TF)-Fe was accumulated in follicular fluid with increasing the follicular diameter, and that TF receptor (TFR1) was localized in granulosa cells of pig. Thus, we hypothesized that TF-Fe would be a factor in the induction of developmental competence of porcine oocytes.
To mimic the follicular development environment, cumulus-oocyte complexes (COCs) were cultured in pre-IVM medium (low dose of FSH) without or with Holo-TF (monoferric or diferric TF) or Apo-TF (non-iron bond TF). After pre-IVM without or with Holo-TF, COCs were cultured in IVM medium (high dose of FSH and EGF) without or with Holo-TF.
Cultivation with Holo-TF increased the expression of follicular development maker ( and ), E2 production, and proliferative activity of cumulus cells, whereas cultivation with Apo-TF did not show these positive effects. The treatment with Holo-TF during pre-IVM, but not during IVM, dramatically induced oocyte maturation with increasing the blastocyst rate.
We succeeded in showing for the first time that the cultivation with Holo-TF in pre-IVM can produce embryos in pig with high efficiency.
由于体外成熟(IVM)后培养的卵母细胞发育能力较低,因此有必要改进IVM方法以实现高效的后代生产。在本研究中,我们发现转铁蛋白(TF)-铁随着卵泡直径的增加而在卵泡液中积累,并且TF受体(TFR1)定位于猪的颗粒细胞中。因此,我们推测TF-铁可能是诱导猪卵母细胞发育能力的一个因素。
为模拟卵泡发育环境,将卵丘-卵母细胞复合体(COCs)在无或有全铁转铁蛋白(单铁或双铁TF)或脱铁转铁蛋白(非铁结合TF)的IVM前培养基(低剂量FSH)中培养。在有或无全铁转铁蛋白的IVM前培养后,将COCs在无或有全铁转铁蛋白的IVM培养基(高剂量FSH和EGF)中培养。
用全铁转铁蛋白培养可增加卵泡发育标志物(和)的表达、E2的产生以及卵丘细胞的增殖活性,而用脱铁转铁蛋白培养则未显示出这些积极作用。在IVM前而非IVM期间用全铁转铁蛋白处理可显著诱导卵母细胞成熟并提高囊胚率。
我们首次成功表明,在IVM前用全铁转铁蛋白培养可高效生产猪胚胎。