مقايسه تعداد اسپرم هاي زنده در نمونه هاي انساني با استفاده از سه روش؛ تورم هيپواسموتيك، V/C Assay و ايوزين- نيگروزين: يك مطالعه آزمايشگاهي

Comparison of Viable Human Sperm Count Using Three Different Methods; Hypo-Osmotic Swelling Test, Viability/Cytotoxicity Assay, and Eosin-Nigrosin Technique: A Laboratory Study

Background and Objectives: Sperm viability is one of the semen parameters that should be noted in male infertility approaches. Depending on the laboratoryʹs facilities and especially the purpose of the experiment, i.e, diagnostic or therapeutic, one or more of the diagnostic techniques are employed. In the present study, three methods; Hypo-Osmotic Swelling Test (HOST), Viability/Cytotoxicity Assay (VCA), and Eosin-Nigrosin Technique (ENT) have been compared in terms of cost, required equipments, diagnostic value, and rapidity. Materials and Methods: In this laboratory study semen samples from six healthy volunteers referred to the Andrology laboratory, Hallamshire hospital, Sheffield, UK were investigated in 2002. In 4 ependorfs, 180~1 of 5 x l O" sperm/ml of prepared sperm using percoll gradient method were decanted. Three of the ependorfs were treated with 20JlI of Chlamydia LPS at fmal concentration of 0.1ug/rnl for 6 h at 37°C in 5% CO2 for 6 h and the last ependorf was treated with 20~1 of 1 x EBSS as control. After 6h incubation the sperm viability was measured using HOST, VCA, and ENT methods. The data was then analysed using t-test and One-way ANOVA. Results: The findings of HOST, VCA, and ENT indicated that sperm mortality rate enhanced markedly in the presence of lipopolysaccharide from Chlamydia trachomatis at 0.1ug/ml; [35.5±4.1% (HOST), 33.8±3.3% (VeA), 34.4±3.9% (ENT), p