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Sperm plasma membrane integrity measurement: a combined method
Author(s) -
Carreras A.,
Ramirez J. P.,
Mendoza C.
Publication year - 2009
Publication title -
andrologia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.633
H-Index - 59
eISSN - 1439-0272
pISSN - 0303-4569
DOI - 10.1111/j.1439-0272.1992.tb02665.x
Subject(s) - propidium iodide , andrology , sperm , staining , biology , chemistry , medicine , apoptosis , biochemistry , genetics , programmed cell death
Summary. Sperm plasma membrane characteristics were measured by a combined method consisting of the hypo‐osmotic swelling test and staining with either the eosine Y (HOS‐eosine test) or propidium iodide dye (HOS‐propidium test). Sperm samples were washed and resuspended in BWW medium (fraction I). Aliquots of the washed spermatozoa were treated by a swim‐up technique to select motile spermatozoa (fraction II). After separation of motile cells, residual sperm pellets were treated separately (fraction III). These three fractions were subjected to the hypo‐osmotic swelling test, lipid peroxidation measurement, and the HOS‐eosine and HOS‐propidium tests. The HOS‐eosine test makes it possible to distinguish 4 types of spermatozoa: type 1: HOS+/eosine ‐; type 2: HOS ‐/ eosine ‐; type 3: HOS ‐/eosine + and type 4: HOS+/eosine+ (Fig. 1). HOS‐propidium test shows equal results as HOS‐eosine test. Fraction I spermatozoa showed 55.2 ± 3.6% type 1; 12.6±1.0 type 2; 28.0±2.9 type 3; and 4.2±0.6 type 4 cells. Fraction II spermatozoa were characterized by high percentages of type 1 cells, low percentages of types 3 and 4, and very low values of lipid peroxidation (5 times smaller than fraction I). Fraction III showed a low percentage of type 1, a high percentages of the other types, and an enhanced value of lipid peroxidation (2 times higher than fraction I). The prognostic value of the HOS‐eosine test was evaluated in an IVF programme. Preliminary results show that a high incidence of types 2 and 4 spermatozoa is often associated with fertilization failure.