Effects of BSA and FCs on in vitro maturation of bitch oocytes at transportation and storage temperatures of +4 Cº and +38 Cº

This study consists of two basic parts. In the first chapter, it is aimed to determine the effects of transport of ovaries obtained by ovariohystectomy method on maturation at transport temperature of +4 ° C and +38.5 ° C. In the second part of the study, it was planned to compare the effects of 0.3% BSA and 5% FCS protein sources, which are added to the medium for in vitro maturation of dog oocytes. One of the ovarian pairs taken from ovario-hysterectomized dogs was brought to the laboratory environment in two different containers containing antibiotics (Gentamicin Sulfate, 50 mg / ml) PBS solution at +4 ° C and the other at +38.5 ° C. After removing the excess oil and tissue around the ovaries, they were washed (+38 ºC) and placed in PBS. Oocytes obtained by slicing method were transferred to washing petrils. In the selection of oocytes to be separated for in vitro maturation, oocyte selections were made considering a solid shingles pellucida, at least 3-4 rows of sandulus cells, homogeneous and dark colored vitellus presence filling the shingles. Within two temperatures, three working groups were prepared at least two hours before, covered with mineral oil (Sigma), added antibiotics (Gentamycin Sulfate, 50 mg / ml) +38 ° C and 5% CO2 in a liincurator environment. transferred to ripening medium (TCM 199 with Hepes Modification, Sigma M-2520). For +4 ºC Transport Temperature; 1st Group (Control): TCM 199 + 2,2 gr / lt NaHCO3 + 0,23 mM Na Pyruvate (pH: 7.3; Osmality: 288 mOsmol), 2nd Group (BSA): TCM199 + 0.3% BSA (Fraction V, Sigma A8806) + 2.2 gr / lt NaHCO3 + 0.23 mM Na Pyruvate (pH: 7.3; Osmality: 288 mOsmol), Group 3 (FCS): TCM199 + 5% FCS (Fetal Calf Serum Biochrom S 0115) + 2.2 gr / lt NaHCO3 + 0.23 mM Na Pyruvate (pH: 7.3; Osmality: 288 mOsmol) For +38.5 ºC Transport Temperature; Group 4 (Control): TCM 199 + 2,2 gr / lt NaHCO3 + 0,23 mM Na Pyruvate (pH: 7.3; Osmality: 288 mOsmol), 5. Group (BSA): TCM199 + 0.3% BSA (Fraction V, Sigma A8806) + 2.2 gr / lt NaHCO3 + 0.23 mM Na Pyruvate (pH: 7.3; Osmality: 288 mOsmol), Group 6 (FCS): TCM199 + 5% FCS (Fetal Calf Serum Biochrom S 0115) + 2.2 g / lt NaHCO3 + 0.23 mM Na Pyruvate (pH: 7.3; Osmality: 288 mOsmol) Then, oocytes were matured for 72 hours in a +38.5 ºC incubator environment where 5% CO2 and 100% humidity was provided in order to reach the MII stage from the Germinal Vesicle (GV) stage. At the end of the maturation period, the sand cells of the oocytes were mechanically removed using vortex. Then it was immersed in 0.7% KCl solution for 4-5 minutes and then it was fixed by squeezing between the coverslip. It was stained with Hoechst 33258 solution. Chi Square test was used to evaluate oocytes. P <0.05 level was accepted to be important in terms of statistical difference. In statistical comparison of those who completed a single (M II) stage between the groups; The BSA group at +4 ° C was determined to be superior to the other control groups than the FCS groups at +4 ° C and +38 ° C (p <0.05In conclusion, in line with the results obtained in the study, it is necessary to add the medium protein in the in vitro maturation studies of dog oocytes; It can be said that 0.3% BSA as protein source as a supplement to maturation is more effective than + 38.5 ° C compared to 5% FCS and transportation temperature at +4 ° C. Keywords: Female dog, Oocyte, In vitro Maturation, BSA, FCS, Transport temperatures