HEPARIN AND HYPOTAURINE SUPPLEMENTATION IMPROVE THE FERTILIZATION RATE OF SHEEP OOCYTES MATURED IN MEDIA CONTAINING L-CARNITINE IN VITRO

R. Rahmatullah, M. Setiadi, I. Supriatna
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Abstract

This study to do examine the effectiveness of using heparin and hypotaurine to improve the in vitro fertilization rate. In first stage, oocytes were matured in the maturation medium supplemented with 0.3 mg/mL L-carnitine to improve nuclear maturation. In second stage, matured oocytes were fertilized with semen incubated in fertilization medium supplemented with 0.1 IU/mL heparin and 10 µM hypotaurine. The final concentration of sperm used was 5x106 spermatozoa/mL, and its interaction with oocytes was carried out for 12 hours in 5% CO2 incubator at 38.5° C. The results showed that percentage of oocytes that reached metaphase II was significantly higher (P0.05) in media supplemented with 0.3 mg/mL L-carnitine compared to control (88.2±4.9% vs 79.8±5.1%). Furthermore, fertilization rate was characterized by formation of 2 PN was significantly higher (P0.05) in media supplemented with heparin and hypotaurine than control (82.6±2.0% vs 73.3±2.4%). In conclusion, L-carnitine supplementation at dose of 0.3 mg/mL on maturation media was effective in increasing percentage of matured oocytes. The combination of 0.1 IU/mL heparin and 10 µM hypotaurine in fertilization medium was also successful in elevating the percentage of fertilized oocytes.
肝素和次牛磺酸的添加提高了体外含左旋肉碱培养基中成熟的绵羊卵母细胞受精率
本研究旨在探讨肝素与次牛磺酸联合应用对提高体外受精率的效果。第一阶段,卵母细胞在添加0.3 mg/mL左旋肉碱的成熟培养基中成熟,以促进核成熟。第二阶段,将成熟卵母细胞与精液在添加0.1 IU/mL肝素和10µM次牛磺酸的受精培养基中孵育。最终使用的精子浓度为5 × 106个/mL,与卵母细胞在38.5℃5% CO2培养箱中相互作用12 h。结果表明,添加0.3 mg/mL左旋肉碱的培养液中达到中期II的卵母细胞比例显著高于对照组(88.2±4.9% vs 79.8±5.1%)(P0.05)。此外,2 PN的形成在添加肝素和次牛磺酸的培养基中显著高于对照组(82.6±2.0% vs 73.3±2.4%)(P0.05)。综上所述,在成熟培养基中添加0.3 mg/mL的左旋肉碱可有效提高成熟卵母细胞的百分率。在受精培养基中加入0.1 IU/mL肝素和10µM次牛磺酸也能成功地提高受精卵的百分率。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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