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Collection and preservation of porcine polar bodies

Published online by Cambridge University Press:  29 January 2010

Wang Gong-Jin ,
Zhou Xiao-Long ,
Tan Xiao-Dong ,
Yu Jian-Ning ,
Xu Xiao-Bo  and
Fan Bi-Qin
Wang Gong-Jin*
Affiliation:
Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China College of Life Science, Nanjing Normal University, Nanjing 210097, China
Zhou Xiao-Long
Affiliation:
College of Life Science, Nanjing Normal University, Nanjing 210097, China
Tan Xiao-Dong
Affiliation:
College of Life Science, Nanjing Normal University, Nanjing 210097, China
Yu Jian-Ning
Affiliation:
Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
Xu Xiao-Bo
Affiliation:
Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
Fan Bi-Qin
Affiliation:
Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
*
*Corresponding author. E-mail: wgjphd@yahoo.cn.
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Abstract

Mature porcine oocytes containing first polar bodies (Pb I) were obtained by in vitro culture of follicle oocytes from ovaries obtained from a local abattoir, and zygotes with second polar bodies (Pb II) were grown after in vitro fertilization of the mature oocytes. Extrusion, biological activity and morphology of Pb I and Pb II were statistically analysed. Polar bodies were isolated and collected from oocytes by enzyme digestion or micromanipulation. Their vigour under different preservation conditions was analysed and evaluated using a Trypan blue staining method. The results showed that 66.7% of the oocytes extruded Pb I after 40 h of in vitro mature culture of oocytes, and 49.7% of the zygotes extruded Pb II 20 h after in vitro fertilization. The efficiency of isolation of Pb II by micromanipulation significantly exceeded that by enzyme digestion, the Pb I and Pb II isolated by micromanipulation presenting with good vigour and normal morphology (95.3% versus 58.9%). The survival rates of Pb I and Pb II were 63.3% and 93.1% for 4 h at 39°C, 85.0% and 72.9% for 40 h at 4°C, and over 95.0% and 84.6% for less than 7 days at −20°C. In comparison with the above preservation conditions for Pb I and Pb II, the results for cryopreservation were best, with rates of survival as high as 89.1% for Pb I and 87.9% for Pb II for preservation periods of over a month, and rates of normal morphology of 97.8% and 95.7%, respectively. The Pb I and Pb II could be isolated and preserved effectively, for use in further research on the recombination of oocytes and zygotes.

Keywords

first polar bodysecond polar bodyisolationpreservation
Type
Research Papers
Information
Chinese Journal of Agricultural Biotechnology , Volume 6 , Issue 3 , December 2009 , pp. 183 - 189
Copyright
Copyright © China Agricultural University 2009

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