Improving outcome of in vitro sperm activation using non–liquefied versus liquefied semen of oligoasthenozoospermic patients
Abstract
Objective: To evaluate the efficacy of in vitro sperm activation (ISA) using non-liquefied versus liquefied asthenozoospermic semen samples for improvement of sperm parameters.
Materials and Methods: Fifty six oligoasthenozoospermic (OA) patients (age range: 22-44 years; mean: 32.089 years) were enrolled in this study. OA patients were classified according to type of infertility. Also, duration of infertility was recorded. Fifty six semen samples were collected, and seminal fluid analyses were done involving macroscopic and microscopic examinations were performed according to WHO methodology. Direct swim–up technique was used to separate the motile spermatozoa from seminal plasma. Minimum Essential Medium Eagle (MEME) enriched with 5% Human serum albumin (HSA) was used. One mL of either non–liquefied or liquefied semen was layered beneath 1 mL of MEME enriched with 5% HSA, and placed for incubation in an air incubator at 37 oC for 30 minutes. Then, one drop (10 μL) from upper layer of culture medium was taken using automatic pipette to be examined under high power field (40 X) for assessment of sperm parameters.
Results: According to type of infertility, infertile patients were classified into patients with either primary infertility (no. 46; 82.15 %) or secondary infertility (no. 10; 17.85 %). In contrast to other parameters, significant (P<0.05) reductions were noticed in the percentages of sperm motility and progressive sperm motility for OA patients with primary infertility as compared to OA patients with secondary infertility. All sperm parameters were significantly (P<0.001) enhanced after in vitro activation of liquefied and non-liquefied semen samples when compared to pre-activation. In the present study, best results were achieved for non-liquefied semen samples as compared to liquefied semen samples.
Conclusion: It was concluded that the outcome of ISA was enhanced in regard to sperm parameters when using non-liquefied semen of OA patients. Furthermore, some components of seminal plasma of OA patients may be have harmful effects on certain sperm functions when in vitro incubated for longer periods. Further study is recommended to investigate the effect of in vitro sperm activation from non-liquefied semen on successful rate of artificial insemination husband.
Shaban SF. Male infertility assessment, diagnosis and treatment. Chapel Hill, NC. University of North Carolina, School of Medicine, USA 1998: 1–19.
Wentz AC. Infertility. In: Novak's Textbook of Gyneco- logy. Jones HW; Wentz AC and Burnett LS (eds.). Williams and Wilkins, Baltimore, USA. 1984: 263–302.
Sharlip ID, Jarow JP, Belker AM, Lipshultz LI, Sigman M, Thomas AJ, et al. Best practice policies for male infertility. Fertil Steril 2002; 77: 873–82.
Purvis K, Christiansen E. Infection in male reproductive tract. Impact, diagnosis and treatment in relation to male infertility. Int J Androl 1993; 16: 1–13.
Henkel R, Defosse K, Weissmann N, Miska W. Assessment of the energy consumption of human spermatozoa related to motility. Reprod Dom Animal 2000; 35: 14–19.
Schill WB, Haidl G. Medical treatment of male infertility. In: Infertility: Male and Female. Lunenfeld B, Insler V. (Eds.), Churchill Livingstone, Edinburgh 1993: 575–622.
Sigman M, Howards SS: Male infertility. In: Campbell's Urology. Walsh PC, Retik AB, Vaughan ED, Wein AJ. (Eds.). 7th Edition. Saunders Company, Philadelphia, USA. 1998; 1287–1330.
Bradsbaw KD, Cbantilis SJ,Carr BR. Diagnostic evaluation and treatment algorithms for the infertile patients. In: Textbook of Reproductive Medicine. Bruce RC, Richard EB. (Eds.). 2nd Edition. Appleton and Lange, USA. 1999; Pp: 533–548.
Ng SC, Bongso A, Sathananthan H, Ratnam SS. Micromanipulation, Its relevance to human in vitro fertilization. Fertil Steril 1990; 53: 203–19.
Micic S, Tulic C, Dotlic,R. Kallikrein therapy of infertile men with varicocele and impaired sperm motility. Andrologia 1990; 22: 179–83.
Francavilla F, Romano R, Santucci R, Poccia C. Effect of sperm morphology and motile sperm count outcome of intrauterine insemination on oligozoospermia and/or Asthenozoospermia. Fertile Steril 1990; 53: 892–7.
Marmar JL . The diagnosis and treatment of male infer- tility in the millennium. Int J Fertil 2001; 46: 116–36.
Baker G, Liu DY, Bourne H. Assessment of the male and preparation of sperm for ARTs. In: Handbook of in vitro fertilization.in: Trounson AO, Gardner DK, (eds.) 2nd edition. CRC Press, USA. 2000: 99–126.
Sepalla, M. The world collaborative report on in vitro fertilization and embryo replacement–Current state of the ART. Ann NY Acad Sci 1985; 442: 558–63.
Ravnik SE, Zarutskie PW, Muller CH. Purification and characterization of a human follicular fluid, lipid transfer protein that stimulates human capacitation. Biol Reprod 1992; 47: 1126–33.
16. Cruz RI, Kemmann E, Brandeis VT, Becker KA, Beck M, Beardsley L, et al. A prospective study of intrauterine insemination of processed sperm from men with oligoasthenospermia in superovulated women. Fertil Steril 1986; 46: 673–7.
World health Organization (WHO). WHO laboratory manual for the examination of human semen and sperm cervical mucus interaction. 4 th ed.; New York; Cambridge University Press.1999: 4–59.
NAFA–ESHRE. Manual on basic semen analysis 2002:1–34.
Fakhrildin MB M–R. The effect of bovine follicular fluid on motility and vitality of human spermatozoa during sperm activation in vitro. Dirasat Journal 1999; 26: 248–55.
Fakhrildin MB M–R. Unsuccessful treatment of severely oligoasthenozoospermic patients using gonadotropins to improve sperm production and motility. Kufa Med Journal. 2006; 9: 410–419.
Fakhrildin MB M–R. Correlation between seminal fluid analysis and levels of prolactin in seminal plasma and serum of healthy men and infertile patients. Kufa Med Journal 2007; 10: 326–34.
Fakhrildin MB M–R. The impact of seminal fluid infections and treatment on sperm functions and fertilizing capacity in infertile patients. Ph.D. Thesis, College of Science, Al–Mustansirya University, Baghdad, IRAQ. 2000; Pp: 86–153.
Kouty, B.K. Results. In: An evaluation of hypo– osmotic swelling test regarding: seminal fluid analysis, sperm preparation techniques, and intrauterine insemination for infertile patients. M.Sc. Thesis, IVF Institute of Embryo Research and Infertility Treatment, Al–Nahrain University, Baghdad, IRAQ. 2007; Pp: 47–97.
Matthews GJ, Goldstein M, Henry JM, Schlegel PN. Non–bacterial pyospermia: a consequence of clomiphene citrate therapy. Int J Fertil Menopausal Stud 1995; 40: 187–91.
Matkov TG, Zenni M, Sandlow J, Levine LA. Preoperative semen analysis as a predictor of seminal improvement following varicocelectomy. Fertile Steril 2001; 75: 63–8.
Bostofte E. Prognostic parameters in predicting pregnancy. ACTA Obstet Gynecol Scand 1987; 66:617–24.
Makler A. Modern methods in semen analysis evaluation. In: Progress in Infertility. Behrman, S.J., Kistner, R.W. and Patton, G.W. (eds.). Little,Brown and Company, Boston, USA. 1988: 633–661.
Matson PL. External quality assessment for semen analysis and sperm antibody detection: results of a pilot scheme. Hum Reprod 1995; 10: 620–5.
Verheyen G, Tournaye H, Staessen C, De Vos A, Vandervorst M, Van Steirteghem A. Controlled comparison of conventional in vitro fertilization and intracytoplasmic sperm injection in patients with Asthenozoospermia. Hum Reprod 1999; 14: 2313–9.
Wikland M, Wik O, Steen Y, Qvist K, Söderlund B, Janson PO. A self–migration method for preparation of sperm for in vitro fertilization. Hum. Reprod. 1987; 2: 191– 5.
Fisch B, Kaplan–Kraicer R, Amit S, Zukerman Z, Ovadia J, Tadir Y. The relationship between sperm parameters and fertilizing capacity in vitro: a predictive role for swim–up migration .J In Vitro Fert Embryo Transf 1990; 7: 38–44.
Steen Y, Wikland M, Janson PO, Wik O. A new method for the treatment of sperm in an IVF and ET program. J IVF–ET 1986; 3: 173– 8.
Rowell P, Braude P. Assisted conception. I—General principles. BMJ 2003; 327: 799–801.
Schlegel PN, Girardi SK. In vitro fertilization for male factor infertility. J Clin Endocrinol Metab1997; 82:709–16.
Good NE, Winget GD, Winter W, Connolly TN, Izawa S, Singh RM. Hydrogen ion buffers for biological research. Biochemistry 1966; 5: 467–77.
King RS, Killian GJ. Purification of bovine estrus– associated protein and localization of binding on sperm. Biol Reprod 1994; 51: 34–42.
Draves, D. Spectrphotometric determination of human serum albumin, quantitative analysis laboratory: A new approach. Department of Chemistry, University of Central Arkansas, USA. 1998; http://chemistry.uca.edu.
Bavister BD, Kinsey DL, Lane M, Gardner DK. Recombinant human albumin supports hamster in vitro fertilization. Hum Reprod 2003; 18: 113– 6.
Henkel RR, Schill WB.Sperm preparation for ART.Reprod Biol Endocrinol 2003; 1: 108–20.
Reiss H. Reproductive medicine. Reiss, H. Oxford University Press, Oxford, UK. 1998; P: 4.
Ranch, D.K. Methods for in vitro fertilization United States Patent: 2005; 4–6, 838, 235. http://www.pharmcast.com/PatentToSubWeb/PatentHo me.htm
Guyton, A.C. and Hall, J.E. Reproductive and hormonal functions of the male (and the pineal gland). In: Textbook of Medical Physiology. Guyton, A.C. and Hall, J.E. (eds.). 9th edition. W.B. Saunders Company, Philadelphia, USA. 1996; Pp: 1003–1016.
Burt CT, Chen B. Biochemical consequences of phosphorylcholines low PK with special relevance to human semen: passive pH regulation. Int J Biochem 1999; 22: 115– 9.
Contemporary management of impotence and infertility. Baltimore (MD): Williams & Wilkins; 1988;3–27.
Gonzales GF, Kortebani G, Mazzolli AB. Hypervis- cosity and hypofunction of the seminal vesicles. Systems Biology in Reproductive Medicine 1993; 30: 63–68.
Tauber PF, Zaneveld LJD. Coagulation and lique- faction of human semen. In: Human semen and fertility regulation in men. Hafez E.S.E. (ed.). The C.V. Mosby Company, USA. 1976: 153–66.
Mortimer D. Sperm preparation methods: Andrology Laboratory Corner. J Androl 2000; 21: 334 – 40.
Henkel R, Bittner J, Weber R, Hüther F, Miska W.Relevance of zinc in human sperm flagella relation to motility. Fertil Steril 1999;71:1138–43.
Faro S. Chlamydia trachomatis: female pelvic infection. Am J Obstet Gynecol 1991; 164: 1767–70.
Chan PJ, Su BC, Tredway DR, Whitney EA, Pang SC, Corselli J,et al. White blood cells in semen affect hyperactivation but not sperm membrane integrity in the head and tail regions. Fertile Steril 1994; 61: 986–9.
Kiefer D, Check JH, Katsoff D. The value of motile density, strict morphology and hypo–osmotic swelling test in in vitro fertilization–embryotransfer. Arch Androl 1996; 37: 57–60.
Kunathikom S, Phophong P, Potjanakorn P, Koanantakul S, Choavaratana R, Suksompong S,et al. Hypo–osmotic swelling test and sperm motility after swim–up and two–layer Percoll gradient preparation of frozen–thawed sperm. Thai J Obstet Gynaecol 1998; 10: 47–51.
Hourani CL, Check JH, Baker AF, Hoover LM, Summers DC, Benfer KM. Cumulus removal and addition of follicular fluid possibly improves pregnancy rates with in vitro fertilization for male factor. Arch Androl 1995;34:47–52.
Tsai YH, Liu J, Jairo GE, Eugene K, Gial C, Baramki TA. Establishment of an optimal hypo–osmotic swelling test by examining single spermatozoa in four different low hypo–osmotic solutions. Hum Reprod 1997; 12: 1111–3.
Suleiman SA, Ali ME, Zaki ZM, el–Malik EM, Nasr MA. Lipid peroxidation and human sperm motility: protective role of vitamin E. J Androl 1996; 17: 530–7.
Momen MN, El–Khodary M, Mostafa T , Yossef HA. Total antioxidant status in infertile males with leukocytospermia. Middle East Fertil Soci 1999; 4: 215–21.
Urata K, Narahara H, Tanaka Y, Egashira T, Takayama F, Miyakawa I. Effect of endotoxin–induced reactive oxygen species on sperm motility. Fertil Steril 2001; 76: 163–6.
Turek PJ. Male infertility. In: Smith's General Urology. Tanagho EA, McAninch JW. (eds.). 15th edition. New York, NY: McGraw–Hill, 2000.750–787.
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| Issue | Vol 4, No 1 (March 2010) | |
| Section | Original Articles | |
| Keywords | ||
| Liquefaction Oligoasthenozoospermia In vitro activation | ||
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