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Submitted to: Journal Of Reproduction, Fertility And Development
Publication Type: Review Article Publication Acceptance Date: 7/3/1995 Publication Date: N/A Citation: N/A Interpretive Summary: Technical Abstract: Recent research on the flow cytometry of sperm for the purpose of predetermining gender of offspring has led to a validated method to separate X- from Y-chromosome bearing sperm for use with in vitro fertilization/embryo transfer, intra-tubal insemination or intra cytoplasmic sperm injection. The basis for the method is the sex chromosome specific marker, DNA, which is present in greater amounts in X- bearing sperm than in Y-bearing sperm of mammals. Sperm are exposed to the vital stain, Hoechst 33342 which binds to the minor groove of the DNA helix. Flow cytometric sorting of the sperm using a laser as the excitation source results in populations of Y- or X-bearing sperm that are 85 to 90% pure. Several hundred offspring have been produced from swine, rabbits, sheep and cattle that confirm the predicted sex. The method is currently being applied to the commercial embryo market. The method is not tlikely to be used in conjunction with standard cattle or swine artificial insemination practice in its current form since only about 400,000 sorted sperm can be produced per hour of sorting. The technology has also been applied to human sperm for use by couples that are at-risk to sex-linked disease expression in their offspring. Populations of human sperm have been sorted with X/Y purities of about 80% as confirmed by DNA probe technology and fluorescence in situ hybridization. |
