This work was carried out to develop a defined in vitro maturation medium for buffalo oocytes that would support their developmental capability after IVF. The medium was based on the use of isobutyl 1-methylexanthin (IBMX) and dibutyryl adenosine monophosphate (dbcAMP) to control the intracellular concentration of cAMP. In the control group, buffalo oocytes were matured in TCM-199+10% fetal calf serum (FCS)+10 g/mL FSH+50 g/ml gentamycin. In the experimental group, oocytes aspiration and selection were performed using TCM-199 Hepes+1 mM IBMX+1 mM dbcAMP+4 mg/mL BSA+50 g/ml gentamycin. Maturation medium was TCM-199+4 mg/mL BSA+50 g/mL gentamycin+0.1 mM dbcAMP (High) or 0.01 mm dbcAMP (Low). After maturation, cumulus expansion was evaluated, and oocytes nuclear morphology was determined. Morphological analysis of the cytoplasm was performed on semi-thin sections of oocytes from each group of maturation. After fertilization, cleavage rate and embryo development to morula and blastocyst stages were assessed on day 2, 5 and 8 of culture, respectively. The results indicated that the degree of cumulus expansion was higher (p<0.01) in the control group than in both concentrations of dbcAMP. Nuclear maturation rates in control and low-dbcAMP groups were higher (p<0.01) than in High-dbcAMP group. Moreover, a high concentration of dbcAMP induced degenerative changes in the cytoplasm of the matured oocytes. Cleavage and blastocyst rates of the Low-dbcAMP group were similar to the control group and higher than the High-dbcAMP group. Our data demonstrate that a defined medium based on the combined use of IBMX and dbcAMP during buffalo oocytes collection and in vitro maturation supports embryo in vitro development to the blastocyst stage and can successfully replace the presence of serum and FSH.

Developmental competence of buffalo oocytes matured in vitro in a defined medium supplemented with dibutyryl cyclic adenosine monophosphate / A.S.S. Abdoon, O.M. Kandil, A.M. Luciano, S. Modina, F. Gandolfi - In: Vyzkum v Chovu Skotu (Cattle Research)[s.l], 2004. - pp. 47-53 (( Intervento presentato al 11. convegno International congress on biotechnology in animal reproduction tenutosi a Czech Republic nel 2004.

Developmental competence of buffalo oocytes matured in vitro in a defined medium supplemented with dibutyryl cyclic adenosine monophosphate

A.M. Luciano;S. Modina
Penultimo
;
F. Gandolfi
Ultimo
2004

Abstract

This work was carried out to develop a defined in vitro maturation medium for buffalo oocytes that would support their developmental capability after IVF. The medium was based on the use of isobutyl 1-methylexanthin (IBMX) and dibutyryl adenosine monophosphate (dbcAMP) to control the intracellular concentration of cAMP. In the control group, buffalo oocytes were matured in TCM-199+10% fetal calf serum (FCS)+10 g/mL FSH+50 g/ml gentamycin. In the experimental group, oocytes aspiration and selection were performed using TCM-199 Hepes+1 mM IBMX+1 mM dbcAMP+4 mg/mL BSA+50 g/ml gentamycin. Maturation medium was TCM-199+4 mg/mL BSA+50 g/mL gentamycin+0.1 mM dbcAMP (High) or 0.01 mm dbcAMP (Low). After maturation, cumulus expansion was evaluated, and oocytes nuclear morphology was determined. Morphological analysis of the cytoplasm was performed on semi-thin sections of oocytes from each group of maturation. After fertilization, cleavage rate and embryo development to morula and blastocyst stages were assessed on day 2, 5 and 8 of culture, respectively. The results indicated that the degree of cumulus expansion was higher (p<0.01) in the control group than in both concentrations of dbcAMP. Nuclear maturation rates in control and low-dbcAMP groups were higher (p<0.01) than in High-dbcAMP group. Moreover, a high concentration of dbcAMP induced degenerative changes in the cytoplasm of the matured oocytes. Cleavage and blastocyst rates of the Low-dbcAMP group were similar to the control group and higher than the High-dbcAMP group. Our data demonstrate that a defined medium based on the combined use of IBMX and dbcAMP during buffalo oocytes collection and in vitro maturation supports embryo in vitro development to the blastocyst stage and can successfully replace the presence of serum and FSH.
blastocyst; c-AMP; culture media; cytoplasm; embryo culture; embryonic development; in vitro fertilization; morphology; oocyte maturation; oocytes
Settore VET/01 - Anatomia degli Animali Domestici
2004
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/184258
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