Momozawa Kenji
School of Veterinary Medicine Kitasato University Towada Aomori Japan.
Reprod Med Biol. 2020 Jul 6;19(4):365-371. doi: 10.1002/rmb2.12337. eCollection 2020 Oct.
In the present study, I evaluated the usefulness of Medium RD, with mixed RPMI1640 and Dulbecco's modified Eagle's medium (1:1, v/v), as a chemically defined medium for in vitro maturation (IVM) of bovine oocytes.
In vitro maturation was performed in 10 mmol/L HEPES-buffered TCM199 (mTCM199), 10 mmol/L HEPES-buffered Medium RD (mRD), and mTCM199 supplemented with fetal bovine serum fraction (mTCM199 + FBS fraction) that served as control. Cumulus-oocyte complexes were matured for 24 hours in three different media supplemented with follicle-stimulating hormone, estradiol-17β, and polyvinylpyrrolidone. Nuclear maturation of oocytes, their developmental competence into blastocysts after in vitro fertilization (IVF) and mitochondrial distribution in oocytes were investigated.
There was no difference in the ratio of matured oocytes regardless of IVM media. The percentage of morula stage was higher in mRD than in mTCM199 group ( < .05) at 120-144 hours after IVF, although the blastocyst rates between groups were not significantly different at 168-216 hours. IVM in mRD increased the percentage of oocytes with diffused mitochondrial distribution compared with the immature and mTCM199 and had similar percentage of oocytes in mTCM199 + FBS fraction.
Medium RD would be useful as a chemically defined medium for IVM of bovine oocytes.
在本研究中，我评估了混合RPMI1640和杜氏改良伊格尔培养基（1:1，v/v）的RD培养基作为牛卵母细胞体外成熟（IVM）的化学成分明确的培养基的实用性。
体外成熟在10 mmol/L HEPES缓冲的TCM199（mTCM199）、10 mmol/L HEPES缓冲的RD培养基（mRD）以及补充了胎牛血清组分的mTCM199（mTCM199 + FBS组分）中进行，后者作为对照。卵丘 - 卵母细胞复合体在补充了促卵泡激素、雌二醇 - 17β和聚乙烯吡咯烷酮的三种不同培养基中成熟24小时。研究了卵母细胞的核成熟、体外受精（IVF）后发育成囊胚的能力以及卵母细胞中的线粒体分布。
无论IVM培养基如何，成熟卵母细胞的比例均无差异。在IVF后120 - 144小时，mRD组的桑葚胚阶段百分比高于mTCM199组（P <.05），尽管在168 - 216小时时各组间的囊胚率无显著差异。与未成熟组和mTCM199相比，mRD中的IVM增加了线粒体分布弥散的卵母细胞百分比，且与mTCM199 + FBS组分中的卵母细胞百分比相似。
RD培养基可作为牛卵母细胞IVM的化学成分明确的培养基。
