[논문]Analysis of Sexed Sperm by Flow Cytometry in Hanwoo (Korean Native Cattle)

Yoo, Han-Jun; Lee, Kyung-Jin; Lee, Yong-Seung; Yoon, Pil-Sang; Park, Joung-Jun; Kim, Hyeong-Cheol; Park, Choon-Keun

Analysis of Sexed Sperm by Flow Cytometry in Hanwoo (Korean Native Cattle) 원문보기

Reproductive & developmental biology = 한국동물번식학회지, v.36 no.1, 2012년, pp.1 - 6

Yoo, Han-Jun (Developmental Biotechnology Laboratory, Myung-poom Hanwoo Consulting) , Lee, Kyung-Jin (College of Animal Life Sciences, Kangwon National University) , Lee, Yong-Seung (College of Animal Life Sciences, Kangwon National University) , Yoon, Pil-Sang (Developmental Biotechnology Laboratory, Myung-poom Hanwoo Consulting) , Park, Joung-Jun (Developmental Biotechnology Laboratory, Myung-poom Hanwoo Consulting) , Kim, Hyeong-Cheol (Hanwoo Experiment Station, National Livestock Research Institute) , Park, Choon-Keun (College of Animal Life Sciences, Kangwon National University)

Abstract ▼ AI-Helper

This study evaluated a sexed sperm ability to produce embryos by flow cytometer. Hanwoo bulls sperm were separated to X and Y sperm via Hoechst 33342 stained with near UV laser or performed the pre-sorted without near UV laser beam in flow cytometry. Pre-sorted sperm had significantly higher viability ($84{\pm}1.15%$, $p$<0.05) compared to other sorted groups in frozen-thawed semen. For fresh semen, pre-sorted sperm had the higher viability ($79{\pm}3%$, $p$<0.05) than those of the X and Y sperm ($44.7{\pm}1.67$ and $41.7{\pm}1.2%$) separated by differences of DNA content. On the other hand, pre-sorted and X sperm sorted according to differences in DNA content had significantly higher viabilities ($24.3{\pm}1.2$ and $25.7{\pm}0.9%$, $p$<0.05) compared to that of the sorted Y sperm ($13.7{\pm}1.2%$) in the hypoosmotic swelling test. The proportion acrosome reaction in the sorted X sperm was higher ($55.0{\pm}1.7$ and $45.0{\pm}1.5%$) than those of the sorted Y-sperm ($32.3{\pm}0.9%$, $p$<0.05). However, the sperm morphologies of the sorted groups were not significantly differences. In conclusion, the sex-sorting procedure by flow cytometry affected some characteristics of Hanwoo sperm. Further study is needed to determine the optimal procedures to enhance male and female embryos and sorting accuracy.

주제어

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문제 정의

Therefore, the objective of the present study was to investigate sperm ability to produce embryos after sexsorting by flow cytometry in Hanwoo (Korean native cattle).

제안 방법

To minimize these limitations, the Percoll gradient and BSA were tested during sperm preparation. Moreover, an additional experiment (data not shown) was conducted to examine the differences between three different collection media: HEPES buffer containing 2 % egg yolk or 10 % seminal plasma, and 0.4 % BSA HEPES buffer containing 2 % egg yolk in order to achieve the highest viability and storability before sperm analysis. Test was performed where egg yolk, BSA and seminal plasma from boar semen were added to the collection tube before or after sperm sorting.
4 % BSA HEPES buffer containing 2 % egg yolk in order to achieve the highest viability and storability before sperm analysis. Test was performed where egg yolk, BSA and seminal plasma from boar semen were added to the collection tube before or after sperm sorting. The Percoll gradient and egg yolk effected viability, and the egg yolk and seminal plasma increased motility after sperm sorting.
The status of the sperm acrosome was assessed using the Coomassie Brilliant blue G-250 dye method (Moller et al., 1990) with slight modifications. Briefly, the dye solution was prepared by suspending 15 mg of Coomassie Brilliant blue G-250 dye in 10 ml of 3.

대상 데이터

The authors are grateful to researcher Joung-Hee Park, Biotechnology Analysis Lab, Central Laboratory Kangwon National University, Korea and gratefully acknowledges the Hoengseong Livestock Cooperative and Institute of Animal Resources of Kangwon National University for providing the semen samples and analysis.

데이터처리

Differences among sperm analysis mean values from the sperm sorting method results were processed using Duncan's multiple range tests.
Statistical analysis was performed with analysis of variance (ANOVA) using SAS (version 9.1, SAS Institute Inc., Cary, NC, USA). Differences among sperm analysis mean values from the sperm sorting method results were processed using Duncan's multiple range tests.

이론/모형

Sperm preparation and staining were based on the method described by Johnson et al. (1989) in order to maintain viability through sorting and fertilization. Briefly, pretreated aliquots of fresh or frozen- thawed semen were stained with 40 μM Hoechst 33342 (Sigma-Aldrich, St.
The hypo-osmotic swelling test (HOST) is based on the semi-permeability of the intact cell membrane, which causes sperm to swell under hypo-osmotic conditions, when an influx of water results in an expansion of cell volume (Drevius and Eriksson, 1966). The test method was introduced by Jeyendran et al. (1984). Briefly, HOST solution (150 mosm/l) was prepared by dissolving 0.

성능/효과

Also, significantly lower viability sorted sperm were analyzed sorted Y sperm (62.7±1.2 %) by differences of DNA content in exceptional control group (p<0.05).
In conclusion, the sex-sorting procedure by flow cytometry affected some characteristics of Hanwoo sperm. Further study is needed to determine the optimal procedures to enhance male and female embryos and sorting accuracy.

후속연구

In conclusion, the sex-sorting procedure by flow cytometry affected some characteristics of Hanwoo sperm. Further study is needed to determine the optimal procedures to enhance male and female embryos and sorting accuracy.

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저자의 다른 논문 :

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