CAJ | 학술논문

[目的]改善猪裸卵的体外成熟质量,为建立稳定有效的裸卵培养体系提供科学依据。[方法]以第一极体排出比例、卵母细胞谷胱甘肽含量、亮甲酚蓝阳性率以及早期胚胎发育潜能等为主要衡量指标研究了卵泡壁颗粒细胞共培养对猪裸卵胞质成熟的影响。[结果]体外成熟结果表明,相对于DO组,DO＋MGC组卵母细胞排除第一极体的比例无显著差异,但核成熟的进程得到了改善,更接近于COC组;成熟卵母细胞GSH含量检测结果表明,DO＋MGC组平均每个卵母细胞谷胱甘肽含量（光密度1053.67）与COC组（1426.00）和COC＋GC组（1541.00）差异不显著,但DO组（724.67）显著低于COC组和COC＋GC组（P〈0.05）;G6PDH活性检测结果表明,DO＋MGC组BCB阳性卵母细胞比例（88.26%）与COC组（92.75%）以及DO组（82.86%）差异均不显著,但DO组显著低于COC组（P〈0.05）;成熟卵母细胞孤雌激活后的发育结果表明,DO＋MGC组卵裂率（94.98%）和囊胚率（43.67%）显著高于DO组卵裂率（52.54%）和囊胚率（8.97%）（P〈0.05）,与COC组卵裂率（97.11%）和囊胚率（38.30%）差异不显著;成熟卵母细胞体外受精后的发育结果表明,DO＋MGC组卵裂率（72.65%）与DO组卵裂率（63.59%）无显著差异,但DO＋MGC组囊胚率（17.79%）显著高于DO组（9.88%）（P〈0.05）。[结论]卵泡壁颗粒细胞共培养可以显著改善裸卵体外成熟的胞质成熟质量并进而提高后续发育潜力。
[목적]개선저라란적체외성숙질량,위건립은정유효적라란배양체계제공과학의거。[방법]이제일겁체배출비례、란모세포곡광감태함량、량갑분람양성솔이급조기배태발육잠능등위주요형량지표연구료란포벽과립세포공배양대저라란포질성숙적영향。[결과]체외성숙결과표명,상대우DO조,DO＋MGC조란모세포배제제일겁체적비례무현저차이,단핵성숙적진정득도료개선,경접근우COC조;성숙란모세포GSH함량검측결과표명,DO＋MGC조평균매개란모세포곡광감태함량（광밀도1053.67）여COC조（1426.00）화COC＋GC조（1541.00）차이불현저,단DO조（724.67）현저저우COC조화COC＋GC조（P〈0.05）;G6PDH활성검측결과표명,DO＋MGC조BCB양성란모세포비례（88.26%）여COC조（92.75%）이급DO조（82.86%）차이균불현저,단DO조현저저우COC조（P〈0.05）;성숙란모세포고자격활후적발육결과표명,DO＋MGC조란렬솔（94.98%）화낭배솔（43.67%）현저고우DO조란렬솔（52.54%）화낭배솔（8.97%）（P〈0.05）,여COC조란렬솔（97.11%）화낭배솔（38.30%）차이불현저;성숙란모세포체외수정후적발육결과표명,DO＋MGC조란렬솔（72.65%）여DO조란렬솔（63.59%）무현저차이,단DO＋MGC조낭배솔（17.79%）현저고우DO조（9.88%）（P〈0.05）。[결론]란포벽과립세포공배양가이현저개선라란체외성숙적포질성숙질량병진이제고후속발육잠력。
[Objective] This study aimed to improve the in vitro maturation quality of denuded porcine oocytes and provide scientific basis for establishing a stable and efficient denuded oocyte culture system. [Method] The first polar body extrusion rate, oocyte glutathione （GSH） content, positive rate of brilliant cresyl blue （BCB） staining and development potential of activated oocytes or fertilized oocytes were employed as main indicators to investigate the effects of follicular mural granulosa cell （MGC） coculture on cytoplasmic maturation of cumulus cell-removal oocytes （Denuded Oocyte, DO）. [Result] According to in vitro maturation results, compared with DO group, the first polar body extrusion rate of porcine oocytes in DO＋MGC group was not significantly different, but the nuclear maturation process was improved and was more similar to that in COC （cumulus-oocyte complex） group. Detection of GSH content in mature oocytes showed that there was no significant difference between DO＋ MGC group （optical density of 1 053.67） and COC group （optical density of 1 426.00） or between DO＋MGC group and COC＋GC group （optical density of 1 541.00）, however, GSH content in mature oocytes of DO group （optical density of 724.67） was significantly lower than that of COC group and COC＋GC group （P0.05）. Detection of glucose-6-phosphate dehydrogenase （G6PDH） activity showed that there was no significant difference in BCB positive oocyte rate between DO ＋MGC group （88.26% ） and COC group （92.75%） or between DO＋MGC group and DO group （82.86% ）, however, BCB positive oocyte rate of DO group was significantly lower than that of COC group （P0.05）. Furthermore, the cleavage rate and blastocyst rate of activated mature oocytes derived from DO ＋MGC group （94.98% and 43.67% , respectively） were significantly higher than those from DO group （52.54% and 8.97%, respectively）（P0.05）, and were not significantly different compared with those from COC group （97.11% and 38.30%, respectively）. In addition, the cleavage rate of fertilized oocytes derived from DO＋MGC group （72.65%） showed no significant difference compared with that from DO group （63.59%）, but the blastocyst rate of DO＋MGC group was significantly higher than that of DO group （9.88%）（P0.05）. [Conclusion] MGC coculture can significantly improve the in vitro cytoplasmic maturation quality of denuded porcine oocytes, thereby enhancing the subsequent developmental potential.