Journal of Reproduction & Infertility

Journal of Reproduction & Infertility

Effect of human sperm MTT viability assay on outcome of intraycytoplasmic sperm injection

Authors
Mohammad Hossein Nasr-Esfahani 1
Roshanak Aboutorabi 2
Ebrahim Esfandiary 2
1 Embryology Department, Royan Institute, Iranian Academic Center for Education, Culture & Research (ACECR), Tehran, Iran; Isfahan Fertility and Infertility Center, Tehran, Iran
2 Anatomy Department, Faculty of Medicine, Isfahan Medical Sciences University, Isfahan, Iran
Abstract
Sperm MTT Viability Assay has been shown to be a suitable test for differentiation of viable from non-viable sperms. In this procedure MTT is converted to observable purple MTT Formazan by mitochondrial dehydrogenase in the midpiece region and therefore viable sperms can be distinguished which makes this test suitable for ICSI. Therefore, in order to study the effect of MTT positive sperms on fertilization, cleavage and blastocyst formation, 109 fresh human oocytes (metaphase II) were divided in to two groups; one group was injected with MTT positive sperms and the other one was taken as control. The results of study showed that there is not significant difference with respect to fertilization, cleavege and blastocyst formation between these two groups. Therefore, if MTT proves to be nether mitogenic nor teratogenic, sperm MTT viability assay might be useful for ICSI in patients with absolute or severe asthenospermia, especially in cases with tail abnormality.
Keywords
Human sperm
ICSI
MTT
Viability assay
Eliasson R., Mossberg B., Camner P., Afzelius BA. The immntile-cilia syndrom. A congenital ciliary abnormality as an etiologic factor in chronic airway infections and male sterility. N Engl J Med. 1977;297:1-6.
Yieh-Loong T., Jiaen L., Jairo E., Garcia E., Eugene K., Campton G., Baramki T.A. Establishment of an optimal hypo-osmotic swelling test by examining single spermatozoa in four different htpo-osmotic solutions. Hum Reprod. 1997;12(5):1111-3.
WHO laboratory manual for the examination of human semen sperm-cervical mucus interaction. 3th Edition, Cambridge University Press. 1992; PP52.
Ahmadi A., Soon-Chye N.G. 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):346-50.
Hossein A.M., Rizk B, Bairk S., Huff C., Thorneycroft I.H. Time course of hypo-osmotic swelling test of human spermatozoa; evidence of ordered transition between swelling subtypes. Hum Reprod.1998;13(6):1578-83.
ابوترابی ر، اسفندیاری ا، نصراصفهانی م ح، مردانی م. یک روش نوین جهت بررسی حیات اسپرم با استفاده از MTT. نشریه پزشکی یاخته، سال 3، شماره 9، 1380، ص 6-1.
Knars-Esfahani M.H., Aboutorabi R., Esfandiari E., Mardani M. Sperm MTT viability assay: a new method for evaluation of human sperm viability. J Assist Reprod Genet. 2002;19(10):475-80.
Mosmann T. Rapid colorimetric assay for cellular growth and survival, Application to proliferation and cytotoxisity assay. J Immunol Method. 1983;65:55-63.
Hansen M.B., Nielsen S.E., and 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: 203-10.
El-Nour A.M., Al Mayman H.A., Jaroudi K.A., Coskun S. Effects of the hypo-osmotic sweelling test on the outcome of intracytoplasmic sperm injection for patients with only nonmotile spermatozoa available for injection; a prospective randomized trial. Fertil Steril. 2001;75(3):480-4.
Tasdemir I., Tasdemir M., Tavukculoglu S. Effect of pentoxifylline on immotile testicular spermatozoa. J Assist Reprod Genet. 1998;15:90-2.
Ramirez J.P., Carreras A., Mendoza C Sperm plasma membrane integrity in fertile and infertile men.Andrologia. 1992;24:141-4.
Journal of Reproduction & Infertility
Volume 3, Issue 4
Autumn 2002
Pages 4-13