THE EFFECTIVITY OF CYSTEAMINE SUPPLEMENTATION ON IMPROVING THE IN VITRO FERTILIZATION RATE OF SHEEP OOCYTES

Dona Astari Nurkarimah, E. M. Kaiin, N. W. Karja, M. Setiadi
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Abstract

The purpose of this study was to evaluate the effectiveness of cysteamine supplementation on the maturation medium and/or in vitro fertilization medium with regards to improving the normal fertilization rate of sheep oocytes, which are characterized by the formation of two pronuclei. Grade A and B oocytes were matured in medium-199 with 0.3% bovine serum albumin (BSA), 10 IU/mL follicle stimulating hormone (FSH), 10 IU/mL human chorionic gonadotrophin (hCG), and 50 μg/mL gentamicin added for 24 hours in a 5% CO2 incubator at 39° C. The treatment group was divided into the following groups: a control group with no cysteamine (P1), a group with 100 μM cysteamine in the maturation medium (P2), a group with 100 μM cysteamine in the fertilization medium (P3), and a group with 100 μM cysteamine both in the maturation and fertilization medium (P4). The fertilization was carried out by incubating sperm-oocytes for 12 hours and then staining them with 1% aceto-orcein to observe the formation of a pronucleus. Normal fertilization rates obtained by each treatment group were 56.5% (P1), 57.1% (P2), 57.8% (P3), and 59.9% (P4) with no significant difference (P0.05) between groups. It was concluded that 100 μM cysteamine supplementation in both the maturation medium and fertilization medium was not able to increase the normal fertilization rate of sheep oocytes.
补充半胱胺对提高绵羊卵母细胞体外受精率的效果
本研究的目的是评价在成熟培养基和(或)体外受精培养基中添加半胱胺对提高绵羊卵母细胞正常受精率的效果。绵羊卵母细胞以形成两个原核为特征。A级和B级卵母细胞在培养基中成熟,添加0.3%牛血清白蛋白(BSA)、10 IU/mL促卵泡激素(FSH)、10 IU/mL人绒毛膜促性腺激素(hCG)和50 μg/mL庆大霉素,于5% CO2培养箱中,39℃培养24小时。不添加半胱胺的对照组(P1)、成熟培养基中添加100 μM半胱胺的组(P2)、受精培养基中添加100 μM半胱胺的组(P3)、成熟和受精培养基中同时添加100 μM半胱胺的组(P4)。精子-卵母细胞孵育12小时后用1%乙酰卵黄蛋白染色,观察原核的形成。各组正常受精率分别为56.5% (P1)、57.1% (P2)、57.8% (P3)和59.9% (P4),各组间差异无统计学意义(P0.05)。综上所述,在成熟培养基和受精培养基中添加100 μM半胱胺均不能提高绵羊卵母细胞的正常受精率。
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