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Collection and preservation of porcine polar bodies

Published online by Cambridge University Press:  29 January 2010

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: [email protected].

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.

Type
Research Papers
Copyright
Copyright © China Agricultural University 2009

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