New Technique, Using a Portable CO2 Incubator, for the Production of In Vitro Fertilized Egyptian Buffalo Embryos.

Abstract

Abstract. Buffalo ovaries were collected from a slaughter house and cumulus oocyte complexes were aspirated from follicles 2-6 mm in diameter. A portable CO2 incubator and a CR1aa medium were used for in vitro production of embryos. In the first experiment, numbers of follicles and oocytes per ovary were counted, and classified according to the appearance of the cumulus cells. In the second experiment, in vitro fertilization and cultivation of buffalo oocytes matured in vitro were carried out using semen from three different bulls, which had different sperm motilities (Bull 1: 20%, Bull 2: 60% and Bull 3: 15%, respectively). In the third experiment, the cleavage and transferable embryos of fertilized oocytes from denuded oocytes, and oocytes with intact cumulus cell layers were compared. The percentages of good, fair and poor quality oocytes were 60.9%, 17.8% and 21.4%, respectively. The rates of cleavage and transferable embryos were significantly higher after incubation with semen from Bull 2 (81% and 40.2%) than those after incubation with semen from Bulls 1 (52% and 13.5%, p<0.05 and p<0.01) and 3 (15.8% and 0%, p<0.01). The rates of cleavage and transferable embryos were significantly higher (p<0.01) for oocytes with an intact cumulus than for oocytes without cumulus cells at fertilization.

Effect of Seasons on Ovarian Activity and Developmental Competence of Buffalo Oocytes and Embryos

“…No. of follicles According to Kandil et al [12], the quality of buffalo oocytes (COCs) was assessed. Depending on their cumulus investment and evenly granulated ooplasm, oocytes were divided into 4 groups under a stereomicroscope [90 x] as follows:…”

“…According to the extent of cumulus-cell development, the cytoplasmic maturation of buffalo oocytes was evaluated and divided into 4 grades (G 0, GI, GII, GIII) [12].…”

Effect of melatonin on developmental competence, mitochondrial distribution, and intensity of fresh and vitrified/thawed in vitro matured buffalo oocytes

“…The COCs were examined under a stereo microscope and rinsed for 3-times in oocyte maturation medium. According to Kandil et al [29], buffalo oocyte quality was assessed. Depending on the cumulus investment and equally granulated ooplasm, there were four groups of COCs under a stereomicroscope (90 x) as follows:…”

“…(M5250, Sigma). The COCs were in vitro matured in an incubator at 38.5°C in a humidified atmosphere with 5% CO2 for 22 h. After 22 h from incubation, the cytoplasmic maturation of oocytes assessed according to the degree of cumulus expansion, according to the extent of cumulus-cell development, the cytoplasmic maturation of buffalo oocytes was evaluated and divided into 4-grades according to Kandil et al [29]: G0: Without expansion. GI: With slight expansion.…”