The preparation and culture of washed human sperm: a comparison of a suite of protein-free media with media containing human serum albumin
Asian Pacific Journal of Reproduction
School of Medical and Health Sciences
To compare two suites of culture media (one with HSA and one protein-free (PF) supplemented with methylcellulose) for washing human sperm in IVF.
Semen samples (n = 41) underwent parallel density gradient preparation using PF or HSA-supplemented culture medium and subsequent yield, survival, morphology and motility were compared.
The PF medium resulted in a significantly higher sperm yield (P < 0.0001), but similar sperm morphology (P = 0.822) and 24-h survival (P = 0.11). There was, however, a lower percentage of progressively motile sperm (P < 0.0001) and a higher proportion of sperm demonstrating non-progressive motility (P < 0.0001) in the PF medium when observed on a Makler Chamber, apparently an artefact as a similar sperm motility index was measured using a Sperm Quality Analyser (P = 0.83). Attachment of sperm in PF medium to the glass chamber reduced with time and any differences had disappeared after 6 min on the counting chamber.
These results support the use of PF media supplemented with methylcellulose as an alternative to HSA, although a modification to the manufacturer's protocol of 6-min pre-incubation before assessing sperm motility must be used. Further studies should investigate the function of such sperm prepared in PF medium.
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Peirce,K.L., Roberts, P., Ali, J., Matson, P.L.(2015). The preparation and culture of washed human sperm: A comparison of a suite of protein-free media with media containing human serum albumin, Asian Pacific Journal of Reproduction. 4(3) 222-227.