Adib Maryam, Seifati Seyed Morteza, Ashkezari Mahmood Dehghani, Khoradmehr Arezoo, Rezaee-Ranjbar-Sardari Roshan, Tahajjodi Somayyeh Sadat, Aflatoonian Behrouz
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.
Int J Reprod Biomed. 2020 Dec 21;18(12):1019-1028. doi: 10.18502/ijrm.v18i12.8023. eCollection 2020 Dec.
To increase the results of infertility treatment, many efforts have been made to improve the treatment methods. As assisted reproductive technology is mainly using cell culture methods, one of the approaches to improve this technology is conditioned medium from different sources. It is desirable to apply maturation (IVM) and use oocytes from normal cycles instead of stimulating ovulation.
To investigate the effect of human cumulus cell condition medium (hCCCM) on the IVM of immature mouse oocytes and morphology.
In this experimental study, 240 germinal vesile oocytes were collected from four-six wk-old mice after 48 hr of 5IU pregnant mare serum gonadotropin (PMSG) injection and cultured in hCCCM (test group, n = 120) and DMEM + 20% FBS (control group, n = 120). The IVM rates and changes in perivitelline space (PVS) and shape were investigated at 8, 16, and 24 hr following the culture. The mature (MII) oocytes were subjected to in vitro fertilization (IVF) and the fertilization rate was assessed in three days.
A significant difference was observed between the maturation rates in the hCCCM and control groups (24.16% vs 0%; p = 0.001), as well as morphologic changes between the two groups (p = 0.04, p = 0.05). The development rate for MII oocytes attained from IVM in the hCCCM group was 27.58% (2-cell) and 6.89% (4-cell). Data displayed that hCCCM is an effective medium for oocytes maturation compared to the control medium.
hCCCM supports oocyte growth and maturation. Moreover, hCCCM changes the oocyte shape and size of perivitelline space.
为提高不孕治疗的效果，人们已做出诸多努力来改进治疗方法。由于辅助生殖技术主要采用细胞培养方法，改进该技术的方法之一是使用不同来源的条件培养基。应用未成熟卵母细胞体外成熟（IVM）并使用正常周期的卵母细胞而非刺激排卵是可取的。
研究人卵丘细胞条件培养基（hCCCM）对未成熟小鼠卵母细胞体外成熟及形态的影响。
在本实验研究中，对4至6周龄小鼠注射5国际单位孕马血清促性腺激素（PMSG）48小时后收集240个生发泡卵母细胞，分别在hCCCM（实验组，n = 120）和DMEM + 20%胎牛血清（对照组，n = 120）中培养。培养8、16和24小时后，研究体外成熟率、卵周隙（PVS）变化及形态变化。将成熟（MII）卵母细胞进行体外受精（IVF），并在三天后评估受精率。
hCCCM组与对照组的成熟率之间存在显著差异（24.16%对0%；p = 0.001），两组之间的形态变化也存在显著差异（p = 0.04，p = 0.05）。hCCCM组通过IVM获得的MII卵母细胞的发育率为27.58%（2细胞）和6.89%（4细胞）。数据显示，与对照培养基相比，hCCCM是一种有效的卵母细胞成熟培养基。
hCCCM支持卵母细胞生长和成熟。此外，hCCCM会改变卵母细胞的形状和卵周隙大小。
