OBJECTIVE: To evaluate the incidence of apoptosis after vitrification warming of mouse ovaries. DESIGN: Experimental study. SETTING: University-based research laboratory. ANIMAL(S): Twelve- to 14-day-old National Medical Research Institute female mice. INTERVENTION(S): Vitrification of mouse ovaries using ethylene glycol. MAIN OUTCOME MEASURE(S): Follicle viability assessment by trypan blue testing, morphologic examination by hematoxylin-eosin staining and transmission electron microscopy, apoptosis assessment using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling method and DNA laddering technique. RESULT(S): No statistically significant difference in follicle viability was observed between vitrified and nonvitrified ovaries. On transmission electron microscopy, vitrified ovaries showed a well-preserved ultrastructure. No sign of apoptosis was observed morphologically or by transferase-mediated deoxyuridine triphosphate nick end-labeling technique in either fresh or vitrified-warmed mouse ovaries. Despite the presence of a laddering pattern of DNA in control induced thymic tissue, no similar pattern was observed in fresh or vitrified-warmed ovaries. CONCLUSION(S): The data suggest that the vitrification technique does not induce apoptosis in mouse ovarian tissue investigated just after warming.
OBJECTIVE: To evaluate the incidence of apoptosis after vitrification warming of mouse ovaries. DESIGN: Experimental study. SETTING: University-based research laboratory. ANIMAL(S): Twelve- to 14-day-old National Medical Research Institute female mice. INTERVENTION(S): Vitrification of mouse ovaries using ethylene glycol. MAIN OUTCOME MEASURE(S): Follicle viability assessment by trypan blue testing, morphologic examination by hematoxylin-eosin staining and transmission electron microscopy, apoptosis assessment using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling method and DNA laddering technique. RESULT(S): No statistically significant difference in follicle viability was observed between vitrified and nonvitrified ovaries. On transmission electron microscopy, vitrified ovaries showed a well-preserved ultrastructure. No sign of apoptosis was observed morphologically or by transferase-mediated deoxyuridine triphosphate nick end-labeling technique in either fresh or vitrified-warmed mouse ovaries. Despite the presence of a laddering pattern of DNA in control induced thymic tissue, no similar pattern was observed in fresh or vitrified-warmed ovaries. CONCLUSION(S): The data suggest that the vitrification technique does not induce apoptosis in mouse ovarian tissue investigated just after warming.
Authors: Fariba Khosravi; Robert L Reid; Ashraf Moini; Farid Abolhassani; Mojtaba R Valojerdi; Frederick W K Kan Journal: J Assist Reprod Genet Date: 2013-10-25 Impact factor: 3.412