Open AccessEnhanced separation of X and Y bearing sperm cells by a combined density gradient centrifugation evaluated by fluorescence in situ hybridization of the Y‐chromosome
Author(s) -
Andersen Claus Yding,
Byskov Anne Grete
Publication year - 1997
Publication title -
acta obstetricia et gynecologica scandinavica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.401
H-Index - 102
eISSN - 1600-0412
pISSN - 0001-6349
DOI - 10.3109/00016349709050068
Subject(s) - percoll , sperm , centrifugation , differential centrifugation , semen , andrology , microbiology and biotechnology , biology , chromatography , chemistry , medicine
Objectives . Fluorescent in situ hybridization of decondensed sperm cells was used to evaluate the effectiveness of separating X‐ and Y chromosome bearing sperm cells using a combination of Percoll and NycoPrep media. Material and methods . Semen samples from seven healthy men were divided into three equal aliquots. One aliquot served as an untreated control, the other two were either applied to a seven‐step Percoll gradient (100%‐40%), or to a seven‐step Percoll gradient (100%‐40%) underlayered with 100% NycoPrep for sperm purification. After centrifugation Y‐bearing sperm cells in the pellet and in the control sample was determined by fluorescent in situ hybridization. Results . A significantly reduction of Y‐bearing sperm cells were observed in the pellet of the Percoll + NycoPrep centrifugation (39.3% Y‐bearing sperm cells) compared to the control sample and to Percoll centrifugation without NycoPrep. The Percoll centrifugation without NycoPrep did not show a significant reduction of Y‐bearing sperm cells in the pellet compared to the control sample. Conclusion . By the addition of a medium with a density higher than 100% Percoll a significantly increased separation of X and Y‐bearing sperm cells can be achieved. This study opens new perspectives for improving separation of X‐ and Y‐bearing spermatozoa by simple centri‐fugations. The described centrifugation technique can be applied in connection with preim‐plantation diagnosis in order to create more pre‐embryos of the desired gender.