Rezaee Ranjbar Sardari Roshan, Dehghani Ashkezari Mahmood, Seifati Seyed Morteza, Izadi Mahin, Golzadeh Jalal, Mazaheri Fahime, Adib Maryam, Aflatoonian Behrouz
Department of Biology, Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran.
Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Cell J. 2023 Jul 25;25(7):455-460. doi: 10.22074/cellj.2023.1987024.1217.
Some reports have indicated that conditioned medium from growing mouse embryonic stem cells (ESCs) provides a supportive condition for small follicles growing, oocyte maturation, and following embryo growth. The aim of this study is assessing in vitro maturation (IVM) and consequent fertilization (IVF) outcome of immature mouse oocytes using human embryonic stem cells conditioned medium (HESCM).
In this experimental study, 240 germinal vesicle (GV) oocytes were took from NMRI female mice, aged 4-6 weeks, 48 hours before injection of 5 IU pregnant mare serum gonadotropin (PMSG). 120 GV oocytes without cumulus cells were cultured in each of the groups. 120 GV were cultured in HESCM as test groups and also 120 GV cultured in human embryonic stem cells medium (HESM) as control groups. After evaluating the metaphase II (MII) oocyte maturation rate at 8, 16 and 24 hours, the MII oocytes subsequently were fertilized and the two-cell embryo development rate was recorded at days 1, 2, and 3. Statistical analysis was performed by using the generalized estimating equations (GEE) method that calculated their rate ratio.
Our data indicated there are significant differences between the maturation rates in HESCM and HESM (P=0.004), also the two-cell embryo development was significant between two culture media (P=0.00).
Similar to some other studies, the secretome of the HESCM showed a significant impact on the IVM outcomes in mice.
一些报告表明，生长中的小鼠胚胎干细胞（ESC）的条件培养基为小卵泡生长、卵母细胞成熟及后续胚胎发育提供了支持条件。本研究的目的是评估使用人胚胎干细胞条件培养基（HESCM）对未成熟小鼠卵母细胞进行体外成熟（IVM）及后续受精（IVF）的结果。
在本实验研究中，从4至6周龄的NMRI雌性小鼠体内获取240个生发泡（GV）期卵母细胞，于注射5国际单位孕马血清促性腺激素（PMSG）前48小时取材。每组培养120个无卵丘细胞的GV期卵母细胞。120个GV期卵母细胞在HESCM中培养作为试验组，另外120个GV期卵母细胞在人胚胎干细胞培养基（HESM）中培养作为对照组。在8、16和24小时评估中期II（MII）期卵母细胞成熟率后，将MII期卵母细胞随后进行受精，并在第1、2和3天记录二细胞胚胎发育率。采用广义估计方程（GEE）方法进行统计分析并计算其率比。
我们的数据表明，HESCM和HESM中的成熟率之间存在显著差异（P = 0.004），两种培养基之间的二细胞胚胎发育也有显著差异（P = 0.00）。
与其他一些研究相似，HESCM的分泌产物对小鼠的IVM结果有显著影响。
