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Updated: Aug 26, 2026
Sperm Collection of Differential Quality Using Density Gradient Centrifugation
Published on: November 29, 2018
Separation of human immunodeficiency virus type 1 from motile sperm by the double tube gradient method versus other
Joseph A Politch1, Chong Xu, Lynne Tucker
1Fearing Research Laboratory, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
Objective:
To compare a new sperm-processing device, utilizing a double tube and gradient, with other commonly used sperm-processing methods for their abilities to exclude human immunodeficiency virus type 1 (HIV-1) from the motile sperm fraction while retaining maximum sperm yield.
Design:
Laboratory experiments.
Setting:
Academic research environment.
Patient(S):
Healthy HIV-1 seronegative men between the ages of 25 and 55.
Intervention(S):
Semen samples were spiked with HIV-1 (MN HIV-1 strain; range of concentrations: 10(-1)-10(6) TCID(50)) and subjected to one of the following sperm-processing methods: double sperm tube with discontinuous gradient of sperm separation medium formed inside (double tube gradient), conventional single tube gradient, swim-up or single tube gradient followed by swim-up (gradient/swim-up), which is the method currently used to minimize HIV-1 in semen used for assisted reproductive technology (ART) procedures. For the gradient techniques, Percoll, ISolate, and PureSperm sperm separation media were compared for efficacy of HIV-1 removal. The amount of HIV-1 remaining in the motile sperm pellet after processing was measured by reverse transcription-polymerase chain reaction (RT-PCR), NucliSens assay, or quantitative HIV-1 culture. A hemacytometer was used to microscopically assess motile sperm count.
Main Outcome Measure(S):
HIV-1 RNA copy number, decrease in HIV-1 TCID(50), motile sperm yield.
Result(S):
The double tube gradient technique was significantly better than all other methods in the removal of HIV-1 from the motile sperm fraction, and produced significantly higher sperm yields in comparison with the gradient/swim-up method.
Conclusion(S):
The double tube gradient technique is a relatively simple and effective method that reduces the risk of infection by HIV-1 while producing good sperm recovery.
