Impact of freezing/thawing technique on sperm DNA integrity in HIV-1 patients

Abstract

Introduction According french legislation, sperm freezing/ thawing procedures are used to prevent ART contaminations in couple with HIV-1 infected men. We determined sperm nuclear fragmentation rate before and after selection and freezing/thawing in HIV-1 14 patients.Methods Two groups of patients were studied: 20 control patients with normal sperm (group 1) and without viral infection and 20 fertile treated HIV-1 patients (group 2). DNA fragmentation was evaluated using terminal uridine nick end labeling, before and after gradient selection, and after cryopreservation and thawing procedures. Results DNA fragmentation rates in fresh semen were increased in HIV patients (6.38% vs 3.39%) (p<0.05) compared with control patients. After sperm migration, fragmentation rates were significantly lower (p<0.0001) in the two groups compared with fresh sperm rates. After freezing/thawing, values were similar to those of fresh semen with an increased rate (p<0.01) for HIV-1 patients, with respectively 3.40% and 5.18% rates in control and infected patients. HIV-1-infected patients treated by antiretroviral therapy showed a significant increase in sperm DNA fragmentation in fresh sperm and also after freezing/ thawing procedures, but these two fragmentation rates were not significantly different. Conclusion So, freezing/thawing procedures do not seem to impair sperm DNA and preserve probability of conception for couples with HIV-1 infected men.

Cryopreservation of Human Spermatozoa Decreases the Number of Motile Normal Spermatozoa, Induces Nuclear Vacuolization and Chromatin Decondensation

“…Cryopreservation was performed as previously described elsewhere (Frainais et al, 2010). Briefly, the sperm suspension was frozen by a trained operator after drop‐wise dilution with a cryoprotectant (Spermfreeze, 15% glycerol in HEPES buffer; Fertipro NV).…”

“…Cryopreservation was found to induce chromatin decondensation (as evaluated by aniline blue staining; Hammadeh et al, 1999; Fortunato et al, 2012), or DNA denaturation (as evaluated by an acridine orange test; Spanò et al, 1999; Gandini et al, 2006; Dejarkom and Kunathikom, 2007), whereas another group failed to see negative effects of cryopreservation on chromatin stability when using 1% sodium dodecyl sulfate + 6 mM EDTA (Huret, 1984). More recently, cryopreservation was shown to increase sperm DNA fragmentation (Donnelly et al, 2001; De Paula et al, 2006; Frainais et al, 2010; Zribi et al, 2010; Gosálvez et al, 2011). However, other studies have not evidenced DNA damage after sperm cryopreservation (Duru et al, 2001; Isachenko et al, 2004).…”

High-magnification sperm selection does not decrease the aneuploidy rate in patients who are heterozygous for reciprocal translocations

“…Liquefied, fresh semen samples were evaluated according to the WHO guidelines [34] and prepared on a two-layer, discontinuous concentration gradient (PureSperm 100, Nidacon, Sweden), as described previously [15]. In a glass-bottomed dish (the WillCo-dish from WillCo Wells BV, The Netherlands), 5-10 μl of the sperm fraction obtained after density gradient selection were diluted into a 4 μl microdroplet of 10 % polyvinylpyrrolidone in flushing medium (FertiPro N.V., Belgium).…”

Sperm FISH analysis of a 44,X,der(Y),t(Y;15)(q12;q10)pat,rob(13;14)(q10;q10)mat complex chromosome rearrangement

“…Next, slides were washed three times in PBS and analysed after counterstaining with DAPI. Spermatozoa with DNA fragmentation fluoresced blue and green, as previously described (Frainais et al ., ). One thousand spermatozoa were counted.…”