Comparison between Two Cryo-devices for Vitrification of Immature Oocytes of Indigenous Zebu Cows in Bangladesh

서지정보

ㆍ발행기관 : 한국수정란이식학회 ㆍ수록지정보 : Journal of Embryo Transfer / 32권 / 4호
ㆍ저자명 : Sk Mohiuddin Choudhury, Mohammad Musharraf Uddin Bhuiyan, Mohammad Moshiur Rahman, Md. Masudur Rahman, Md. Newaz Sharif, Jayonta Bhattacharjee, Farida Yeasmin Bari, Nasrin Sultana Juyena

목차

INTRODUCTION
MATERIALS and METHODS
1. Chemicals and media
2. Collection of ovaries
3. Collection of oocytes
4. Selection of occytes for culture
5. Vitrification and warming
6. In vitro culture of oocytes for maturation
7. Evaluation of oocytes for maturation
8. Staining of oocyte
9. Experimental approaches and statistical analysis
RESULTS
DISCUSSION
ACKNOWLEDGEMENT
REFERENCES

영어 초록

Cryopreservation of oocytes by vitrification technique may contribute a lot in the field of reproductive biotechnology. The objectives of the present study were to evaluate the effectiveness of two cryo-devices for vitrification of immature oocytes of indigenous zebu cows. Slaughter house derived immature cumulus-oocyte-complexes (COCs) of cows were vitrified using 15% dimethyl sulphoxide (DMSO) as cryoprotective agent (CPA) with 0.5 mol sucrose in TCM 199 supplemented with 20% FBS. Vitrification of COCs was completed after immediate plunging of COCs loaded cryotop or French mini straw into the liquid nitrogen (LN2). Then the COCs containing cryotop or French mini straws were warmed in 0.25 mol sucrose and 20% FBS supplemented TCM 199 followed by in vitro culture in 50 μl droplets of bicarbonate buffered TCM 199 supplemented with 10% FBS, pyruvate, FSH and oestradiol for 24 hrs at 39°C with 5% CO2 in humidified air. After maturation culture, oocytes were denuded and examined under inverted microscope for presence of polar body as the indication of maturation. Denuded oocytes were also stained by whole mount technique using 1% orcein to examine the maturation by presence of MII chromosomes. The in vitro maturation rate was significantly (p<0.05) higher in oocytes vitrified and warmed using crytop (47.1±6.9%) than that of French mini straw (15.9±12.5%). Moreover, in vitro maturation rate was significantly (p<0.05) highe r in control oocytes (not vitrified) (84.5±14.2%) than that of vitrified oocytes. In conclusion, cryotop is better than French mini straw as cryo-device for vitrification of bovine immature oocytes

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