To the Editor:
We read the article “Selection of viable spermatozoa from testicular biopsies: a comparative study between pentoxifylline and hypoosmotic swelling test” by Mangoli et al. (1) with great interest.
For ICSI, in order to achieve fertilization, we need to select and inject live sperm. For ejaculated sperm, the best marker for viability has been motility. However, this study used fresh testicular sperm for intracytoplasmic sperm injection (ICSI). In our experience, fresh testicular sperm are always viable, whether or not they are moving. This is because the testes can only produce live cells, and freshly extracted and processed sperm from the testes will always be alive. In our experience, it is not necessary to wait for fresh testicular sperm to acquire motility in vitro; and fertilization rates with fresh testicular immotile sperm are as high as with fresh testicular motile sperm. This has been documented by other workers as well (2).
The described tests – hypoosmotic swelling test (HOS) and pentoxifylline – may be useful when working with ejaculated sperm, epididymal sperm or with frozen testicular sperm because in these cases the immotile sperm may be dead and therefore cannot be used for ICSI. However, there is no need to subject fresh testicular immotile sperm to viability testing, as this wastes time and can even reduce pregnancy rates (as sperm subjected to HOS can be difficult to use for ICSI, as noted in the study!).
Anjali Malpani, MD
Aniruddha Malpani, MD
Malpani Infertility Clinic
1. Mangoli V, Mangoli R, Dandekar S, Suri K, Desai S. Selection of viable spermatozoa from testicular biopsies: a comparative study between pentoxifylline and hypoosmotic swelling test. Fertil Steril. 2011 Feb;95(2):631-4.
2. Konc J, Kanyó K, Cseh S. The effect of condition/state of testicular spermatozoa injected to the outcome of TESE-ICSI-ET cycles. Eur J Obstet Gynecol Reprod Biol. 2008 Nov;141(1):39-43.
Published online in Fertility and Sterility doi:10.1016/j.fertnstert.2011.03.053
The Authors Respond:
In response to the letter by Dr Anjali Malpani and Dr Aniruddha Malpani on our article regarding use of pentoxifylline to assess viability of testicular sperm, we would like to comment as submitted herewith.
Though it is assumed that testicular sperms are alive when retrieved, their ‘viability duration’ varies from few minutes to 48 hours (1) Therefore it becomes necessary to evaluate viability of the sperm used for ICSI. Hence most andrologists and embryologists depend upon motility as a distinguishing criterion for viability (2-4). Otherwise, all the sperms obtained from testicular biopsy would have fertilized all mature oocytes.
Furthermore, it is documented that fertilization with nonmotile testicular sperm results in higher incidence of mitotic spindle defects (5). Some andrologist even prefer to do testicular sperm extraction (TESE) one day prior in order to observe motility over period of intervals. Authors who published data using testicular sperms preferred motile spermatozoa for obvious reasons, and tried to locate viable sperm either by ‘mechanical touch technique’ (6), by HOS method (7), or using laser beam (8).
The concern of ‘waste of time’ is inappropriate and on the contrary it saves time to select viable sperm by exposing for merely 10 to 15 minutes. It is also well documented that pentoxifylline has no adverse effect on sperm either at the cellular or the nuclear level (9). The comment ‘can even reduce pregnancy rates’ mentioned in relation to HOS in fact is supporting our observation.
Vijay Mangoli, Ph.D.
Fertility Clinic and IVF Centre
1. Kovacic B, Vlaisavljevic V, Reljic M. J Androl. 2006;27:45–52.
2. Bin Wu, Journal of Assisted Reproduction and Genetics; Volume 22, Numbers 11-12, 389-94.
3. Kahraman S, TasdemirM, Tasdemir I, Vicdan K, Ozgur S, Polat G Islk AZ, Biberoglu K, Vanderzwalmen P, Nijs M ;Pregnancies achieved with testicular and ejaculated spermatozoa in combination with intracytoplasmic sperm injection in men with totally or initially immotile spermatozoa in the ejaculate. Hum Reprod;1996; 11, 1343-6
4. Hassan Sallam, Human Reproduction Volume20, Issue12 )/ (Tournaye et al., 1994; Devroey et al., 1996) Pp. 3435-3440.
5. Tasdemir I , Tasdemir M, and Tavukcuoglu S; Effect of Pentoxifylline on immotile testicular spermatozoa. J Assist Reprod Genet;1998;15, 90-92.
6. de Oliveira NM, Vaca Sanchez R, Rodriguez Fiesta S, Lopez Salgado T, Rodriguez R, Bethencourt JC, et al. Pregnancy with frozen thawed and fresh testicular biopsy after motile and immotile sperm microinjection, using the mechanical touch technique to assess viability. Hum Reprod. 2004; 11: 1343-6.
7. Abdel-Fattah Agameya Ashraf Farrag, Yehia El-Garem, Fathy Ezzeldin, Hassan N. Sallam .The Use of the modified Hypo-Osmotic Swelling Test for the Selection of Immotile Testicular Spermatozoa in Patients Treated With ICSI – A Randomized Controlled Study, Fertil Steril. 2006; 85:S8.
8. Aktan AM, Montag M, Duman S, Gorkemli H, Rink K and Yurdakul T Use of a laser to detect viable but immotile spermatozoa. Andrologia. 2004; 36: 366-369
9. Jacob Rajfer, MD. Rev Urol. 2006 Spring; 8(2): 88.
Published online in Fertility and Sterility doi:10.1016/j.fertnstert.2011.03.052