Abstract
The present study aimed to evaluate the effect of supplementing epidermal growth factor EGF , insulin-like growth factor-I IGF-I , fetal bovine serum FBS , and bovine serum albumin BSA in different combinations in a maturation medium MM comprising TCM-199 + 5 µg/mL 17-β estradiol + 5 µg/mL ovine-luteinizing hormone oLH + 5 µg/mL pure follicle-stimulating hormone pFSH + 100 µM/mL cysteamine containing the best concentration of leptin among 10/20/25/30 ng/mL on in vitro maturation IVM and subsequent vitrification of matured goat oocytes. The IVM of oocytes was carried out in a CO2 incubator for 27 h at 38.5 °C. The percentage of oocytes with cumulus expansion and polar body extrusion was found to be highest in 25 ng/mL leptin at 47.37% and 15.63%; rates of 66.67% and 28.57% were obtained when supplemented with 50 ng/mL IGF-I + 10 ng/mL EGF in medium containing 25 ng/mL leptin as compared to the other media. The in vitro matured oocytes were vitrified in 5 M ethylene glycol + 5 M propylene glycol with 0.5 M sucrose in TCM-199 + 20% FBS using liquid nitrogen. The highest percentage of morphologically intact in vitro matured oocytes following vitrification was obtained in medium containing 25 ng/mL leptin 89.29% . It was concluded that addition of IGF-I + EGF with 25 ng/mL leptin in MM yielded a higher rate of maturation of oocytes, and supplementation of 25 ng/mL leptin in MM resulted in a higher rate of morphologically intact vitrified oocytes, indicating the efficiency of leptin for cryotolerance.
Highlights
Goat is an important livestock species in developing countries because of its ability to utilize different types of forages, and it is very versatile for producers of milk, meat, and skin
The rate of in vitro maturation (IVM) obtained with 25 ng/ mL leptin was the highest (67.50%) and significantly (P < 0.01) higher than that obtained with 10, 20, and 30 ng/ mL leptin based on cumulus cell expansion
IVM in 25 ng/mL leptin with Fetal bovine serum (FBS), bovine serum albumin (BSA), and combination The rates of IVM of oocytes based on cumulus expansion and polar body extrusion in different media, M I, M
Summary
Goat is an important livestock species in developing countries because of its ability to utilize different types of forages, and it is very versatile for producers of milk, meat, and skin. Sustained efforts have been made to accelerate genetic gain in goats through the utilization of artificial insemination and multiple ovulation embryo transfer techniques. The culture media employed for in vitro maturation (IVM) affect the proportion of mammalian oocytes undergoing fertilization and influence the subsequent cleavage and developmental competency [1,2]. Varying effects of different additives in culture media were reported for IVM of oocytes. The addition of growth factors, e.g., epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I), stimulated oocyte maturation and had beneficial effects on blastocyst production rates in several species [3]. Fetal bovine serum (FBS) is a common supplement for in vitro and ex vivo
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