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Sperm MTT Viability Assay: A New Method for Evaluation of Human Sperm Viability

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Abstract

Purpose : MTT (3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay is commonly used as a cell proliferation assay. The objective of this study was to evaluate the capability of MTT assay to discriminate between viable and nonviable sperms and compare its efficiency with E&N (eosin and nigrosin) and HOST (hypo-osmotic swelling test).

Methods : MTT assay was modified to obtain optimal result for assessment of sperm viability. After standardization of method, MTT, E&N, and HOST were carried out simultaneously on 57 semen samples from patients referring to Isfahan Fertility and Infertility Center. The correlation coefficient between these tests and sperm motility was calculated using the SPSS statistical program. Specificity and sensitivity of each test was also obtained.

Results : The optimal conditions for sperm MTT viability assay were 2 h after addition of sperm to MTT in HAM'S F10 + 25 mM HEPES + 10% HSA at 37°C and pH 7.4–7.45. Inter- and intra-assay coefficients of variations were 9 and 7%, respectively. The sensitivity and specificity for sperm MTT viability assay, E&N, and HOST were 97, 98, and 99%, and 100, 100, and 83% respectively. High significant correlations were obtained between sperm MTT viability assay, E&N, HOST and motility.

Conclusions : Sperm MTT viability assay can be used as a diagnostic test for discrimination of viable sperms from sperm population.

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REFERENCES

  1. Palermo G, Joris H, Devroey P,Van Steirteghem AC: Pregnancies after intracytoplasmic injection of a single spermatozoon into an oocyte. Lancet 1992;340:8-17

    Google Scholar 

  2. WHO Laboratory Manual for the Examination of Human Semen and Sperm Cervical Mucus Interaction, 3rd edn, Cambridge University Press, 1992

  3. Tsai YL, Liu J, Garcia JE, Katz E, Compton G, Baramki TA: Establishment of an optimal hypo-osmotic swelling test by examining single spermatozoa in four different hypo-osmotic solutions. Hum Reprod 1997;12(5):1111-1113

    Google Scholar 

  4. Hossain AM, Rizk B, Barik S, Huff C, Thorneycroft IH: Time course of hypo-osmotic swellings of human spermatozoa: Evidence of ordered transition between swelling subtypes. Hum Reprod 1998;13(6):1578-1583

    Google Scholar 

  5. Ahmadi A, Ng SC: The single sperm curling test, a modified hypo-osmotic swelling test, as a potential technique for selection of viable sperm for intracytoplasmic sperm injection. Fertil Steril 1997;68(2):346-350

    Google Scholar 

  6. Mosmann T: Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63

    Google Scholar 

  7. Hansen MB, Nielsen SE, Berg K: Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Immunol Methods 1989;119:203-210

    Google Scholar 

  8. Liu J, Tsai YL, Katz E, Compton G, Garcia JE, Baramki TA: High fertilization rate obtained after intracytoplasmic sperm injection with 100% nonmotile spermatozoa selected by using a simple modified hypo-osmotic swelling test. Fertil Steril 1997;68(2):373-376

    Google Scholar 

  9. Mortimer D: Technician Training and Quality Control Aspect in Practical Laboratory Andrology, Oxford University Press, New York, 1994

    Google Scholar 

  10. Ahmadi A, Ng SC: Developmental capacity of damaged spermatozoa. Hum Reprod 1999;14(9):2279-2285

    Google Scholar 

  11. Eliasson R, Mossberg B, Camner P, Afzelius BA: The immotilecilia syndrome:Acongenital ciliary abnormality as an etiologic factor in chronic airway infections and male sterility. N Engl J Med 1977;297:1-6

    Google Scholar 

  12. Tasdemir I, Tasdemir M, Tavukcuoglu S: Effect of Pentoxifyline on immotile testicular spermatozoa. J Assist Reprod Genet 1998;15:90-92

    Google Scholar 

  13. Sallam HN, Farrag A, Amaya AF, Ezzeldin F, Eid A, Sallam A: The use of a modified hypo-osmotic swelling test for the selection of viable ejaculated and testicular immotile spermatozoa in ICSI. Reproduction 2001;16(2):272-276

    Google Scholar 

  14. Ramirez JP, Carreras A, Mendoza C: Sperm plasma membrane integrity in fertile and infertile men. Andrologia 1992;24:141-144

    Google Scholar 

  15. Jeyendran RS, Van derVen HH, Perez-Pelaez M: Development of an assay to assess the functional integrity of the human sperm membranes and its relationship to other semen characteristics. J Reprod Fertil 1984;70:219-228

    Google Scholar 

  16. Casper RF, Meriano JS, Jarvi KA, Cowan L, Lucato MN: The hypo-osmotic swelling test for selection of viable sperm for intracytoplasmic sperm injection in men with complete asthenospermia. Fertil Steril 1996;65:972-976

    Google Scholar 

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Authors and Affiliations

  1. Department of Clinical and Experimental Embryology and Andrology, Royan Institute, Tehran, Iran

    Mohammad Hossein Nasr-Esfahani

  2. Isfahan Fertility and Infertility Center, Isfahan, Iran

    Mohammad Hossein Nasr-Esfahani

  3. Department of Anatomy, Isfahan University of Medical Sciences, Isfahan, Iran

    Roshanak Aboutorabi, Ebrahim Esfandiari & Mohammad Mardani

Authors
  1. Mohammad Hossein Nasr-Esfahani
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  2. Roshanak Aboutorabi
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  3. Ebrahim Esfandiari
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  4. Mohammad Mardani
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Correspondence to Mohammad Hossein Nasr-Esfahani.

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Nasr-Esfahani, M.H., Aboutorabi, R., Esfandiari, E. et al. Sperm MTT Viability Assay: A New Method for Evaluation of Human Sperm Viability. J Assist Reprod Genet 19, 477–482 (2002). https://doi.org/10.1023/A:1020310503143

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