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The Use of Styrofoam Box for Chikso (Korean Brindled Cattle) Semen Cryopreservation with Liquid Nitrogen

칡소 동결 정액 생산을 위한 스티로폼상자와 액체질소 이용 방법

  • Kim, Sung Woo (Animal Genetic Resources Research Center, National Institute of Animal Science, RDA) ;
  • Ko, Yeoung-Gyu (Animal Genetic Resources Research Center, National Institute of Animal Science, RDA) ;
  • Lee, Jae-Yeong (Animal Genetic Resources Research Center, National Institute of Animal Science, RDA) ;
  • Kim, Chan-Lan (Animal Genetic Resources Research Center, National Institute of Animal Science, RDA) ;
  • Hwang, In-Sul (Animal Biotechnology Division, National Institute of Animal Science, RDA)
  • 김성우 (농촌진흥청 국립축산과학원 가축유전자원센터) ;
  • 고응규 (농촌진흥청 국립축산과학원 가축유전자원센터) ;
  • 이재영 (농촌진흥청 국립축산과학원 가축유전자원센터) ;
  • 김찬란 (농촌진흥청 국립축산과학원 가축유전자원센터) ;
  • 황인설 (농촌진흥청 국립축산과학원 동물바이오공학과)
  • Received : 2019.12.16
  • Accepted : 2020.04.03
  • Published : 2020.04.29

Abstract

A styrofoam box is used as a simple and easy freezing method to preserve animal semen as a livestock genetic source. This study optimized the methods of freezing chikso brindled cattle semen. To test the freezing box, the motility of spermatozoa was compared between two box sizes (length×width×heigh) with the dimensions of 23.5×30.5×22.5 cm and 25.5×46.5×26.5 cm. The motility of thawed sperm from brindled Korean bulls was used to confirm the efficiency of the freezing boxes. The box having a larger inner space with larger horizontal and height measurements supported better motility after thawing (60.4±5.3% vs 67.2±3.1%) with 10 min of exposure time in liquid nitrogen vapor. The optimized freezing space is estimated to be an essential element for successful freezing results and the larger box could be used for production of more than 60 frozen semen straws. These properties are also helpful to optimize the cryopreservation techniques that would control the quality and quantity of semen straws according to different animal species.

가축유전자원으로서 동결정액을 생산하는 가장 쉬운 방법은 스티로폼박스를 이용한 간이동결법으로 알려져 있다. 본 연구에서는 스티로폼 동결박스를 제작하여 가축의 동결정액 생산에 활용하는 방법을 검토하였으며 칡소 동결정액 생산을 최적화 할 수 있는 방법을 제시하고자 박스의 크기, 액체질소와 노출된 정액 스트로와 거리, 노출시간 및 생산량을 검토하였다. 2가지 동결박스를 비교하여 자료를 확보하였으며 내부 크기는 세로×가로×높이가 23.5×30.5×22.5 cm와 25.5×46.5×26.5 cm로 측정되었다. 액체질소를 5cm 높이로 채우고 액체질소 위 2, 5 및 8cm 높이에서 동결하여 융해 후 생존성을 조사하였다. 칡소 정액을 동결할 경우, 액체질소와의 노출시간은 모두 10분이 적절하였으며 25.5×46.5×26.5 cm 크기의 상자가 높은 생존율을 보여주었다(60.4±5.3% 대 67.2±3.1%). 동결 상자의 최적화 공간은 정자 동결에 가장 중요한 요소로 판단되며 1회 동결 시 최대 생산 가능한 칡소 동결정액은 60개 이상으로 증가시킬 수 있었다. 이러한 정보를 활용하면, 축종에 따라 동결 정액 생산량 결정하고 목적에 맞는 용기를 활용하여 효율적인 동결정액 생산이 가능할 것으로 판단된다.

Keywords

Acknowledgement

Supported by : 농촌진흥청

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