I have tried several different extenders (from phisiological saline,PBS, via different types of TRIS to HTF), but for all of them the motility was not good (
"Motility of rat sperm was highest when stored at 22 °C; motility was 28% and 14% at 72 h in TL-HEPES and PBS extenders, respectively.The motility and membrane integrity of rat sperm fell significantly within 24 h at 4 and 37 °C" (Varisli et al., 2013). Obviously temperature plays a role.
Hi Sylwia, are you adding bovine serum albumin to your media? If not, try adding 0.3% (w/v) BSA to either PBS or a tris buffered diluent. Alternatively, you could use polyvinylalcohol (PVA), which basically does the same thing as BSA, typically stops sperm sticking to each other and improves motility. If neither of these avenues work, you could try a TALP type fertilisation medium instead.
Thank you both for your answers.
@Taylor Pini: PBS and TRIS were without BSA, but HTF (Human Tubular Fluid) I used with Serum Substitute Supplement, so it should have worked. But I can try PBS with BSA or focus on the temperature, as Beatrice recommended.
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