Ju Byung Hyun, Kim You Jin, Park Youn Bae, Kim Byeong Ho, Kim Min Kyu
Division of Animal and Dairy Science, College of Agriculture and Life Science, Chungnam National University, Daejeon 34134, Korea.
MK biotech Inc., Daejeon 34134, Korea.
J Anim Sci Technol. 2024 Sep;66(5):936-948. doi: 10.5187/jast.2024.e68. Epub 2024 Sep 30.
The Conical 9 well dish (C9 well dish) is characterized by a decreasing cross-sectional area towards the base. This design was hypothesized to enhance embryonic development by emulating the physical environment through density modulation. Comparative analyses revealed no significant difference in nuclear maturation rates between the C9 well dish and the 5-well dish. Reactive oxygen species (ROS) generation was lower in the C9 well dish compared to the 5-well dish; however, this difference was not statistically significant. On the second day of culture, the cleavage rate in the C9 well dish was 4.66% higher, although not statistically significant, and the rates of blastocyst development were similar across both dishes. No significant differences were observed in the intracellular levels of glutathione (GSH) and ROS, as well as in the total cell number within the blastocysts between the dish types. The expression of mitogen-related factors, TGFα and IGF-1, in the blastocysts was consistent between the dishes. However, PDGFβ expression was significantly lower in the C9 well dish compared to the 35 mm petri dish. Similarly, the expression of the apoptosis factor Bax/Bcl2l2 showed no significant differences between the two dishes. Despite the marked difference in PDGFβ expression, its impact on blastocyst formation appeared negligible. The study also confirmed the feasibility of culturing a small number of oocytes per donor, collected via Ovum Pick-Up (OPU), with reduced volumes of culture medium and mineral oil, thus offering economic advantages. In conclusion, the present study indicates that the C9 well dish is effective for development of a small quantity of oocytes and embryos, presenting it as a viable alternative to traditional cell culture dishes.
锥形9孔培养皿（C9孔培养皿）的特点是朝着底部横截面积逐渐减小。该设计的假设是通过密度调制模拟物理环境来促进胚胎发育。比较分析显示，C9孔培养皿和5孔培养皿之间的核成熟率没有显著差异。与5孔培养皿相比，C9孔培养皿中的活性氧（ROS）生成较低；然而，这种差异在统计学上并不显著。在培养的第二天，C9孔培养皿中的卵裂率高出4.66%，尽管在统计学上不显著，并且两个培养皿中的囊胚发育率相似。在不同培养皿类型之间，谷胱甘肽（GSH）和ROS的细胞内水平以及囊胚内的总细胞数均未观察到显著差异。囊胚中有丝分裂相关因子TGFα和IGF-1的表达在不同培养皿之间是一致的。然而，与35毫米培养皿相比，C9孔培养皿中PDGFβ的表达显著较低。同样，凋亡因子Bax/Bcl2l2的表达在两个培养皿之间没有显著差异。尽管PDGFβ表达存在明显差异，但其对囊胚形成的影响似乎可以忽略不计。该研究还证实了通过采卵（OPU）收集的每个供体少量卵母细胞在减少培养基和矿物油体积的情况下进行培养的可行性，从而具有经济优势。总之，本研究表明C9孔培养皿对少量卵母细胞和胚胎的发育是有效的，是传统细胞培养皿的一个可行替代方案。
