Effect of antifreeze protein supplementation in vitrification medium on mouse oocyte developmental competence

• Published in: Fertility and sterility Vol. 96; no. 5; pp. 1239 - 1245
• Main Authors: Jo, Jun Woo, M.S, Jee, Byung Chul, M.D, Lee, Jung Ryeol, M.D, Suh, Chang Suk, M.D
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier Inc 01-11-2011; Elsevier
• Subjects: adenosine triphosphate; Adenosine Triphosphate - metabolism; Animals; Antifreeze protein; antifreeze proteins; Antifreeze Proteins - pharmacology; Biological and medical sciences; blastocyst; Blastocyst - drug effects; Blastocyst - metabolism; Blastocyst - pathology; caspases; Cell Membrane - drug effects; Cell Membrane - metabolism; Cell Membrane - pathology; chilling injury; chromosomes; Cryopreservation; Cryoprotective Agents - pharmacology; Diseases of the digestive system; Embryo Culture Techniques; embryogenesis; Female; Fertilization in Vitro; gene expression; Gene Expression Regulation, Developmental - drug effects; Gynecology. Andrology. Obstetrics; Internal Medicine; Medical sciences; metaphase; Mice; Obstetrics and Gynecology; oocyte; oocytes; Oocytes - drug effects; Oocytes - metabolism; Oocytes - pathology; protective effect; protein supplements; Radiotherapy. Instrumental treatment. Physiotherapy. Reeducation. Rehabilitation, orthophony, crenotherapy. Diet therapy and various other treatments (general aspects); spindle; Vitrification; oocyte; Antifreeze protein; spindle; vitrification; Gynecology; Vitrification; Rodentia; Obstetrics; Protein; Vertebrata; Mammalia; Diet therapy; Mouse; Animal; Supplementation; Technique; Oocyte
• ISSN: 0015-0282; 1556-5653
• DOI: 10.1016/j.fertnstert.2011.08.023

Objective To investigate the effect of antifreeze protein (AFP) supplementation during mouse oocyte vitrification on the survival, fertilization and embryonic development. Design Animal study. Setting University laboratory. Animal(s) BDF-1 mice. Intervention(s) In vivo–matured metaphase II oocytes were vitrified with the use of CryoTop by two-step exposure to equilibrium and vitrification solution supplemented or not with 500 ng/mL AFP III. Main Outcome Measure(s) Postwarming survival, fertilization, embryonic development up to blastocyst in vitro, morphology of spindle and chromosome, membrane integrity, adenosine triphosphate (ATP) contents, and several gene expressions. Result(s) In the AFP-treated group, blastocyst formation rate was significantly higher and blastomere count with positive caspase was significantly lower compared with the nontreated group. Rate of intact spindle/chromosome, stable membrane, and ATP contents were significantly higher in AFP group. AFP group showed higher Mad2 and lower Eg5 gene expression. Both vitrification groups showed increased Hsf1 , Zar1, and Zp1 / Zp2 expression and decreased Hook1 and Zp3 expression compared with fresh control samples. Conclusion(s) Supplementation of AFP in vitrification medium has a protective effect on mouse oocytes for chilling injury; it can preserve spindle/membrane integrity and intracellular ATP contents. More stable spindle integrity in the AFP group may be associated with higher Mad2 and lower Eg5 gene expression.