Center for Reproductive Medicine - Innovations in Male Infertility Research

INNOVATIONS IN MALE INFERTILITY RESEARCH

Cleveland Clinic's Center for Reproductive Medicine
1993 - 2008

Semen Banking in Cancer Patients

Sperm cryopreservation has become a complementary element in the therapeutic management in young men with malignant diseases, such as testicular cancer or Hodgkin's disease. A substantial number of these patients, however, present with poor semen quality at semen banking. Controversy surrounds the question of whether sperm cryopreservation in patients with malignant diseases is justified. Although the post-thaw semen quality in cancer patients is poor, it is not clear whether the type of cancer is related to cryopreservation results.

We have examined both type of cancer and response to treatment and demonstrated that these factors are related to pre-freeze or post-thaw semen quality. We also have demonstrated that prediction of post-thaw sperm motility from pre-freeze motility is possible. Pre-freeze or post-thaw semen quality in cancer patients is not affected by the type of disease, with the exception of the pre-freeze motile sperm count in testicular cancer patients. Pre-freeze motility can predict post-thaw motility. The impact of cryopreservation on semen quality appears to be independent of the type of malignancy. The clinical diagnosis of cancer is not an adequate predictor of post-thaw semen quality. Cryopreservation of semen should be offered to cancer patients regardless of the type of disease.

Extensive studies have been performed at our center examining various aspects of cancer and sperm quality after cryopreservation such as: effect of cancer on semen quality after cryopreservation of sperm; value of clinical diagnosis in predicting the quality of cryopreserved sperm from cancer patients; DNA damage in patients with untreated cancer as measured by the sperm chromatin structure assay; investigation of fertilizing capacity of cryopreserved spermatozoa from patients with cancer; why cancer patients request disposal of cryopreserved semen specimens post-therapy; characteristics of cryopreserved semen from men with lymphoma; detection of testicular cancer in men presenting with infertility; and the question of whether semen cryopreservation should be offered to men with testicular cancer.

PDF File (agradoc06.pdf 272 Kb)
Agarwal A, and Newton RA (1991):
The effect of cancer on semen quality after cryopreservation of sperm.
Andrologia 23:329-332.
PDF File (agradoc019.pdf 652 Kb)
Agarwal A, Sidhu RS, Shekarriz M, Thomas AJ Jr (1995):
Optimum abstinence time for cryopreservation of semen in cancer patients.
J Urol 154:86-88.
PDF File (agradoc020.pdf 1026 Kb)
Shekarriz M, Tolentino MV Jr, Ayzman I, Lee JC, Thomas AJ Jr, Agarwal A (1995):
Cryopreservation and semen quality in patients with Hodgkin's disease.
Cancer 75:2732-2736.
PDF File (agradoc24.pdf 601 Kb)
Agarwal A, Tolentino MV Jr, Sidhu RK, Ayzman I, Lee JC, Thomas AJ Jr, Shekarriz M (1995):
Effect of cryopreservation on semen quality in patients with testicular cancer.
Urology 46:382-389.
PDF File (agradoc025.pdf 1195 Kb)
Agarwal A, Shekarriz M, Sidhu RK, Thomas AJ Jr (1996):
Value of clinical diagnosis in predicting the quality of cryopreserved sperm from cancer patients.
J Urol 155:934-938.
PDF File (agradoc031.pdf 860 Kb)
Sharma RK, Kohn S, Padron OF, Agarwal A (1997):
Effect of artificial stimulants on cryopreserved spermatozoa from cancer patients.
J Urol 157:521-524.
PDF File (agradoc32.pdf 490 Kb)
Padron OF, Sharma RK, Thomas AJ Jr, Agarwal A (1997):
Effects of cancer on spermatozoa quality after cryopreservation - a 12 year experience.
Fertil Steril 67:326-331.
PDF File (agradoc39.pdf 337 Kb)
Wang Y, Sharma RK, Agarwal A (1997):
Effect of lipid peroxidation on cryopreserved semen quality in patients with testicular or nontesticular cancer.
Urology 50:414-417.
PDF File (agradoc45.pdf 353 Kb)
Sidhu RS, Wang Y, Agarwal A (1997):
Creatine kinase level and lipid peroxidation rate in human spermatozoa from patients with cancer.
J Assist Reprod Genet 14:538-542.
PDF File (agradoc50.pdf 26 Kb)
Hallak J, Hendin BN, Thomas AJ Jr, Agarwal A (1998):
Investigation of fertilizing capacity of cryopreserved spermatozoa from patients with cancer.
J Urol 159:1217-1220.
PDF File (agradoc51.pdf 54 Kb)
Hallak J, Sharma RK, Thomas AJ Jr, Agarwal A (1998):
Why cancer patients request disposal of cryopreserved semen specimens post-therapy: a retrospective study.
Fertil Steril 69:889-893.
PDF File (agradoc55.pdf 22 Kb)
Schover LR, Agarwal A, Thomas AJ Jr (1998):
Cryopreservation of gameter in young cancer patients [Review].
J Pediat Hematol/Oncol 20:426-428.
PDF File (agradoc63.pdf 69 Kb)
Hallak J, Kolettis PN, Sekhon VS, Thomas AJ Jr, aAgarwal A (1999):
Cryopreservation of sperm from patients with leukemia: is it worth the effort?
Cancer 85:1973-1978.
PDF File (agradoc74.pdf 52 Kb)
Hallak J, Kolettis P, Sekhon VS, Thomas AJ Jr, Agarwal A (1999):
Sperm cryopreservation in patients with testicular cancer.
Urology 54:894-899.

PDF File (agradoc82.pdf 32 Kb)
Hallak J,. Mahran A, Chae J, Agarwal A (2000):
The effects of cryopreservation on semen from men with sarcoma or carcinoma.
J Assist Reprod Genet 17:218-221.
PDF File (agradoc83.pdf 26 Kb)
Agarwal A (2000):
Semen banking in patients with cancer : 20 years experience.
Int J Androl 23:16-19.
PDF File (agradoc87.pdf 50 Kb)
Hallak J, Mahran A, Chae J, Agarwal A (2000):
Poor semen quality from patients with malignancies does not rule out sperm banking.
Urol Res 28:281-284.
PDF File (agradoc89.pdf 31 Kb)
Hallak J, Mahran A.M, Agarwal A (2000):
Characteristics of cryopreserved semen from men with lymphoma.
J Assist Reprod Genet 17:103-107.
PDF File (agradoc90.pdf 29 Kb)
Pasqualotto FF, Agarwal A (2001):
Should we offer semen cryopreservation to men with testicular cancer?
CCF J Med 68:101-102.
PDF File (agradoc92.pdf 118 Kb)
Kobayashi H, Larson K, Sharma RK, Nelson DR, Evenson DP, Toma H, Thomas AJ Jr, Agarwal A (2001):
DNA damage in patients with untreated cancer as measured by the sperm chromatin structure assay.
Fertil Steril 75:469-475.
PDF File (agradoc104.pdf 37 Kb)
Ranganathan P, Mahran AM, Hallak J, Agarwal A (2002):
Sperm cryopreservation for men with nonmalignant, systemic diseases: a descriptive study.
J Androl 23:71-75.
PDF File (agradoc130.pdf 61 Kb)
Pasqualotto FF, Pasqualotto EB, Agarwal A, Thomas AJ Jr. (2003):
Detection of testicular cancer in men presenting with infertility.
Rev Hosp Clin Fac Med S Paulo 58:75-80.
PDF File (agradoc133.pdf 231 Kb)
Agarwal A, Ranganathan P, Kattal N, Pasqualotto FF, Hallak J, Khayal S, Mascha E. (2004):
Fertility after cancer: a prospective review of assisted reproductive outcome with banked semen specimens.
Fertil Steril 81:342-348.
PDF File (agradoc147.pdf 40 Kb)
Nallella KP, Sharma RK, Said TM, Agarwal A (2004):
Inter-sample variability in post-thaw human spermatozoa.
Cryobiology 49:195-9.
PDF File (agradoc156.pdf 429 Kb)
Agarwal A and Said TM (2004):
Implications of systemic malignancies on human fertility. Review Article.
Reprod Biomed Online 6:673-9.
PDF File (agradoc175.pdf 60 Kb)
Agarwal A and Allamaneni SSR (2005):
Disruption of spermatogenesis by the cancer disease process.
J Natl Cancer Inst Monogr 34:9-12.
PDF File (agradoc210.pdf 716 Kb)
Agarwal A and Said TM (2006):
Impaired spermatogenesis in patients treated for hodgkin's lymphoma.
Leukemia & Lymphoma 14: 1-7.
Agarwal A, Fnu, D. (2008):
Technical and ethical challenges of gamete cryopreservation in young cancer patients. Review article.
RBM Online. (In Press)

Molecular Sperm Preparation Protocols

Schematic of separation of cells by MACS

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A. Magnetic-activated cell separation

The externalization of the phospholipid phosphatidylserine from the inner to the outer leaflet of the plasma membrane is a feature of the terminal phase of apoptosis and can be monitored by annexin V binding. In magnetic-activated cell separation (MACS), colloidal super-paramagnetic microbeads bind to annexin V, label the dead and apoptotic spermatozoa and retain them within an external strong magnetic field provided by separation columns. This technique is rapid, non-invasive and allows the selection of apoptotic and non-apoptotic sperm for use in assisted reproductive techology procedures.

PDF File (agradoc172.pdf 640 Kb)
Said TM, Grunewald S, Paasch U, Glander H-J, Baumann T, Kriegel C, Li L, Agarwal A (2005):
Advantage of combining magnetic cell separation with sperm preparation techniques.
Reprod Biomed Online 10:740-6.
PDF File (agradoc173.pdf 179 Kb)
Grunewald S, Paasch U, Said TM, Sharma RK, Glander H-J, Agarwal A (2005):
Caspase activation in human spermatozoa in response to physiological and pathological stimuli.
Fertil Steril 83:1106-12.
PDF File (agradoc177.pdf 149 Kb)
Said TM, Grunewald S, Paasch U, Rasch M, Agarwal A, Glander H-J (2005):
Effects of magnetic-activated cell sorting on sperm motility and cryosurvival rates.
Fertil Steril 83:1442-6.
PDF File (agradoc204.pdf 254 Kb)
Said TM, Agarwal A, Grunewald S, Rasch M, Baumann T, Kriegel C, Li L, Glander H-J, Thomas AJ Jr, Paasch U (2006):
Selection of non-apoptotic spermatozoa as a new tool for enhancing assisted reproduction outcomes: an in-vitro model.
Biol Reprod 74:530-7.
PDF File (agradoc208.pdf 191 Kb)
Grunewald S, Paasch U, Said TM, Rasch M, Agarwal A, Glander H-J (2006):
Magnetic-activated cell sorting before cryopreservation preserves mitochondrial integrity in human spermatozoa.
Cell Tissue Bank 7:99-104.

PDF File (agradoc225.pdf 255 Kb)
Said TM, Agarwal A, Grunewald S, Rasch M, Glander H-J, Paasch U (2006):
Evaluation of sperm recovery following annexin V MACS separation. Review Article.
Reprod BioMed Online 13:336-339.
Paasch U, Agarwal A, Gupta AK, Sharma RK, Grunewald S, Thomas AJ Jr, Glander H-J (2008):
Super-paramagnetic depletion of ejaculated human spermatozoa with apoptotic changes.
Anals NYAS (In Press).
Said T, Agarwal A, Glander JH, Grunewald S, Zborowski M, Paasch U (2008):
Utility of magnetic cell separation as a molecular sperm separation technique.
J Androl (In Press).
PDF File (agradoc240.pdf 224 Kb)
Grunewald S, Said T, Paasch U, Baumann T, Agarwal A, Glander H-J. (2007):
Relationship between sperm apoptosis signaling and oocyte penetration capacity.
Int J of Androl. Jun 15 (Epub ahead of print)

PARP homologues in immature and mature spermatozoa

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Analysis of PARP proteins

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B. Utility of poly(ADP-ribose) polymerase (PARP) in selecting sperm

Poly(ADP-ribose) polymerase (PARP), a nuclear enzyme, plays an important part in repairing damaged DNA. We have for the first time examined its role in ejaculated spermatozoa. PARP homologues have been identified in mature and immature sperm. PARPs have an active role in sperm cell physiology and preventing apoptosis. We also have shown the presence of cleaved PARP in ejaculated human spermatozoa. PARP inhibitors may play different roles in chemical- and oxidative stress-induced sperm damage.

Jha R, Agarwal A, Mahfouz R, Paasch P, Grunewald S, Sabanegh E, Yadav SP, Sharma R (2008):
Determination of Poly(ADP-ribose) polymerase (PARP) homologues in human ejaculated sperm and its correlation with sperm maturation.
Fertil Steril (in press)

Metabolomic Profiling and Oxidative Stress

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Metabolomics is the systematic study of metabolites. Small molecule biomarkers representing functional pathways in a cell, tissue or organism can be important indicators of disturbances in the concentration of various metabolites. Metabolic profiling allows the detection and management of various medical conditions, including male factor infertility. This rapid non-invasive analysis allows the measurement of concentrations of important oxidative stress biomarkers (-CH, -NH -OH and -ROH). Differences in the concentrations of these biomarkers have been shown to have a unique association with seminal plasma of healthy men compared with that from patients with idiopathic infertility. This technique shows promise in assisted reproductive techniques and may have important association between biomarkers of oxidative stress and pregnancy outcome of oocytes and embryos.

PDF File (uronewsv_15_2.pdf 283 Kb)
Agarwal A (2006):
Assessing oxidative stress levels in semen using spectroscopy-based metabolomic profiling- Implications in male infertility.
Urology News, Vol. 15, 2006-2007.

Effects of Cell Phone Use on Sperm

Cell phones emit electromagnetic waves (EMW) of different frequencies that have been linked to adverse effects in human beings. In the United States, cell phones operate at frequencies of 900 -1900 MHz; in most other parts of world cell phones work at 900 -1800 MHz frequencies. Preliminary reports suggest that EMW can reduce the fertilizing potential of spermatozoa and cause sperm DNA damage. Some of the possible mechanisms by which EMW can alter reproductive function are a non-thermal EMW-specific effect, a thermal molecular effect, or a combination of these mechanisms. While some authors have found little or no effect with the use of electronic devices on reproductive function, recent research by our group has shown damaging effects of these devices on semen variables in a study involving 361 men. We currently are assessing the effects of EMW emitted by cell phones (900 - 1900 MHz) on various markers of sperm quality such as count, motility, morphology, viability etc. We will correlate the semen characteristics and exposure to electromagnetic waves from cell phones adjusted by such co-variates as patient demographics, daily environment and lifestyle.

PDF File (agradoc250.pdf 388 Kb)
Fnu D, Makker K, Agarwal A (2007):
Cell phone and infertility: Dissecting the relationship. Review Article.
RBM Online 15:266-70.

Agarwal A, Deepinder F, Sharma RK, Ranga G, Li J (2007):
Effect of cell phone usage on semen analysis in men attending infertility clinic: an observational study.
Fertil Steril. 89:124-8.