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KMID : 1005220120270010037
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Journal of Embryo Transfer
2012 Volume.27 No. 1 p.37 ~ p.43
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Evaluation of Sperm Sex-Sorting Method using Flow Cytometry in Hanwoo (Korean Native Cattle)
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Yoo Han-Jun
Lee Kyung-Jin
Lee Yong-Seung
Lee Chang-Woo
Park Joung-Jun
Cheong Hee-Tae
Yang Boo-Keun
Park Choon-Keun
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Abstract
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This study evaluated a method of sorting X and Y chromosomes based on size using the forward angle light scatter related refractive index (FSC) of a flow cytometer. Hanwoo bulls sperm were separated to X and Y chromosomes by the parameters of FSC or Hoechst 33342 intensity. As a result, using monitor program linked flow cytometry during sorting processing, the purities were 97 ± 0.57 or 96 ± 0.67% for the X-fraction and 96 ± 0.33 or 97 ± 1.33% for the Y-fraction in the two sperm sorting methods. There were no differences in the X and Y ratios (X and Y %) bet- ween the sperm sorting methods based on FSC or DNA content. The proportions of female and male embryos used for in vitro fertilization and development were 66.03 ± 3.31 or 69.37 ± 1.41%, and 70.56 ± 2.42 or 56.11 ± 3.09% when sperm were processed using the sex sorting method by FSC or Hoechst 33342. In conclusion, further study is needed to determine the optimum procedure and improve the nozzle to enhancing sorting accuracy or efficiency. Also, the fin- dings of this study do not negate the possibility that the difference method of sperm sorting cannot use a UV laser beam.
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KEYWORD
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flow cytometry , sperm sorting , FSC , sheath fluid buffer , Hanwoo
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