Department of Obstetrics and Gynecology, Selcuk University, Konya, Turkey.
Department of Reproduction and Artificial Insemination, Selcuk University, Konya, Turkey.
Anim Biotechnol. 2023 Dec;34(8):3887-3896. doi: 10.1080/10495398.2023.2236660. Epub 2023 Jul 19.
This study aimed to investigate the effect of putrescine supplementation to maturation medium during embryo production in cattle on maturation and embryo development/quality. Oocytes obtained from the ovaries of Holstein cattle were used in the study. Obtained cumulus-oocyte complexes were evaluated according to morphological structure, cytoplasmic features, and cumulus cell number, and only Category-I ones were used in the study. Before the maturation step, oocytes were randomly divided into two groups. In the first group (Putrescine group, = 159), 0.5 mM putrescine was added to the maturation medium before maturation. No addition was applied to the maturation medium of the second group (Control group, = 149). Cumulus expansion degrees of oocytes following maturation (Grade I: poor, Grade II: partial, and Grade III: complete) were determined. In addition, the meiosis of oocytes after maturation was evaluated by differential staining. Then the oocytes were left for fertilization with sperm and finally, possible zygotes were transferred to the culture medium. After determining the developmental stages and quality of the embryos after culture, only the embryos at the blastocyst stage were stained with the differential staining method to determine the cell numbers. When the cumulus expansion degrees of the groups were evaluated, the Grade III cumulus expansion rate in the putrescine group was higher than the control group (74.21% and 60.4%; respectively) and the Grade I expansion rate (11.95% and 26.17%; respectively) was found lower ( < .05). When the resumption of meiosis was evaluated according to the cumulus expansion degrees, it was determined that the rate of resumption of meiosis increased as the cumulus expansion increased. In addition, the cleavage rates of oocytes and reaching the blastocyst in the putrescine group were found to be higher than in the control group ( < .05). Moreover, inner cell mass, trophectoderm cells, and total cell counts were found to be higher in blastocysts obtained after the putrescine supplementation to the maturation medium compared to the control group ( < .05). As a result, it was determined that the putrescine supplementation to the maturation medium during embryo production in cattle increased the degree of cumulus expansion and the rate of resumption of meiosis. In addition, putrescine supplementation was thought to increase the rate of reaching the blastocyst of oocytes due to better cell development in embryos.
本研究旨在探讨在牛胚胎生产过程中向成熟培养基中添加腐胺对成熟和胚胎发育/质量的影响。该研究使用来自荷斯坦奶牛卵巢获得的卵母细胞。根据形态结构、细胞质特征和卵丘细胞数量对获得的卵丘-卵母细胞复合物进行评估,仅使用 I 类复合物进行研究。在成熟步骤之前,将卵母细胞随机分为两组。在第一组(腐胺组,n=159)中,在成熟前向成熟培养基中添加 0.5mM 腐胺。第二组(对照组,n=149)的成熟培养基中未添加任何物质。测定成熟后卵母细胞的卵丘扩张程度(I 级:差,II 级:部分,III 级:完全)。此外,通过差异染色评估成熟后卵母细胞的减数分裂。然后,让卵母细胞与精子受精,最后将可能的受精卵转移到培养基中。在培养后确定胚胎的发育阶段和质量后,仅用差异染色法对囊胚期胚胎进行染色以确定细胞数量。当评估两组卵丘扩张程度时,腐胺组的 III 级卵丘扩张率高于对照组(分别为 74.21%和 60.4%),I 级扩张率(分别为 11.95%和 26.17%)较低( < .05)。根据卵丘扩张程度评估减数分裂恢复情况时,发现随着卵丘扩张的增加,减数分裂的恢复率增加。此外,腐胺组的卵母细胞卵裂率和囊胚形成率均高于对照组( < .05)。此外,与对照组相比,在成熟培养基中添加腐胺获得的囊胚中内细胞团、滋养外胚层细胞和总细胞数更高( < .05)。因此,确定在牛胚胎生产过程中向成熟培养基中添加腐胺增加了卵丘扩张程度和减数分裂恢复率。此外,由于胚胎中细胞发育更好,添加腐胺被认为可以提高卵母细胞到达囊胚的速度。