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Effect of Ovarian Storage Temperature and Time on Post Thaw Viability and Maturation Rate of Vitrified Immature Oocytes in Sheep

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BACKGROUND: Vitrification of oocytes as a method of cryopreservation is quite successful, although it is still being standardized because of structural and molecular sensitivity of oocytes to the cooling and freezing process. OBJECTIVE: To investigate the effect of ovarian storage temperature and time on post thaw viability and maturation rate of vitrified immature oocytes in sheep. MATERIALS AND METHODS: The work consisted of oocyte collection from ovaries of abattoir sheep stored at various temperature (0 °C , 4 °C and 25 °C) and time (0 h, 6 h, 12 h and 24 h) combinations and post thaw viability and in vitro maturation rate evaluation. Vitrification was done in 30% vitrification solution, using ethylene glycol and DMSO, with post vitrification evaluation after 1 week 's storage. RESULTS: Significantly higher post thaw viability was observed after storage at 0 °C for 6 h (95.3%) followed by 12 h (85%), with lowest value at 24 h (66.7%). However at 4 °C and 25 °C, values were nonsignificantly higher after 6 h (96.5 and 100% respectively) followed by 12 h (93 and 100%), with significantly lower values after 24 h (85.7 and 90.7%). At storage temperatures of 25 °C and 4 °C, a significantly higher percentage of mature oocytes was observed after 6 h (40 and 39.1%), 12 h (37.3 and 38.1%) and 24 h (34.6 and 36.4%) storage times compared to that at 0 °C (20.3% at 6 h, 14.2% at 12 h and only 13.8% at 24 h). However, at all storage temperatures, there was a tendency for the level of mature oocytes to decrease with storage time, and the levels were significantly lower than the control. CONCLUSION : Acceptable post thaw viability and in vitro maturation rates for oocytes is maintained up to 24 h in ovaries stored at 4 °C and 25 °C compared to at 0 °C, and these conditions may be used for the storage of ovaries meant for oocyte preservation.

Keywords: MATURATION; TEMPERATURE; TIME; VIABILITY; VITRIFICATION

Document Type: Research Article

Affiliations: 1: Assistant Professor, Division of Veterinary Clinical Complex, F.V.Sc and A.H, Shuhama, SKUASTKashmir, Srinagar, India 2: JRF, Division of Veterinary Clinical Complex, F.V.Sc and A.H, Shuhama, SKUAST-Kashmir, Srinagar, India

Publication date: September 1, 2022

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  • CryoLetters is a bimonthly international journal for low temperature sciences, including cryobiology, cryopreservation or vitrification of cells and tissues, chemical and physical aspects of freezing and drying, and studies involving ecology of cold environments, and cold adaptation

    The journal publishes original research reports, authoritative reviews, technical developments and commissioned book reviews of studies of the effects produced by low temperatures on a wide variety of scientific and technical processes, or those involving low temperature techniques in the investigation of physical, chemical, biological and ecological problems.

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