Semen cryobanking for men with cancer--criteria change.

OBJECTIVE: To summarize pregnancies and births after the use of pretherapy cryobanked semen from men with cancer and to reassess the clinical role of semen cryobanking for these patients. DESIGN: Survey of pregnancies and births that have occurred after the use of cryobanked semen from pretherapy cancer patients. SETTING: Survey of the literature and of nine semen banks. PATIENTS: Men with testicular cancer, Hodgkin's diseases, non-Hodgkin's lymphoma, and other types of cancer who cryobanked their semen specimens before therapy. OUTCOME MEASURES: Pregnancies and births resulting from the use of cryobanked semen after artificial insemination by husband (AIH) or other assisted reproductive technology (ART) procedures. RESULTS: The use of AIH and other ART procedures have resulted in 117 documented pregnancies and 115 livebirths. CONCLUSIONS: Cryobanking of semen should be offered to all men diagnosed with cancer because such a procedure provides the only reasonable chance of establishing a pregnancy after therapy that is detrimental to fertility potential. Existing criteria for pretherapy semen cryobanking, therefore, should be revised in view of successful pregnancies even with the use of semen with low spermatozoal densities and motilities, as well as other realized clinical efficacies of ART. Conceptions have occurred after in vitro fertilization (IVF) with < 1 x 10(6) motile spermatozoa. Semen cryobanking should be offered by the attending physician as a viable option for any pretherapy male patient who has any motile sperm and considers the future possibility of having children.

Cryosurvival of Trichomonas vaginalis during cryopreservation of human semen.

Despite a 90% cryosurvival of Trichomonas vaginalis in their growth medium trypticase yeast maltose (TYM) with DMSO, none of these parasites have previously been observed to survive during cryopreservation of infected human semen with glycerol (Andrologia 18, 323 (1986)). This could have been due to the failure of the culture method used to detect low numbers of survivors. The prospects of possible transmission of T. vaginalis by artificial insemination with cryobanked (-196 degrees C) semen prompted an investigation of the cryosurvival of this parasite in the presence of semen with the cryoprotectant glycerol, using a more sensitive culture method for viability evaluation. Semen and seminal fluid from the same 23 ejaculates, as well as culture medium, were inoculated with small clinical numbers of T. vaginalis and evaluated as to their survival before and after cryopreservation. Results indicated: (i) The highest cryosurvival of T. vaginalis (4.5%) was in cryobanked (glycerolated) semen, (ii) semen, as well as glycerol, shows cryoprotective action, and (iii) glycerol reduced survival of parasites in semen, seminal fluid, and TYM medium during exposure prior to freezing. Clinical information on infectivity of small numbers of T. vaginalis and the data presented here suggests that these organisms could be transmitted by artificial insemination with infected cryobanked human semen.

Difference in effect of dog seminal fluid and human seminal fluid and semen on in vitro survival of Trichomonas vaginalis.

The authors examined the ability of dog seminal fluid to serve as an animal model in order to study survival of Trichomonas vaginalis in human urogenital secretions. Two strains were used: SVI-1, a recent isolate from a male patient, and ATCC 30001, a strain that has been cultured for many years. The authors incubated the T. vaginalis strains in dog seminal fluid, human seminal fluid, and human semen, at 37 degrees C for 6, 12, and 24 hrs. For both strains, survival in dog seminal fluid was much poorer than that in human semen and seminal fluid, zero survival being noted at 24 hrs. The SVI-1 strain survived significantly better than ATCC 30001 in all secretions. Data from experiments using isolates from two female patients, but incubated in human seminal fluid alone, agreed with results obtained with SVI-1. Dog seminal fluid may not be a suitable substitute for human secretions in T. vaginalis studies, and long-term culture may decrease the ability of T. vaginalis to survive in the male urogenital tract.

Survival of Trichomonas vaginalis in human semen.

Although exposure of Trichomonas vaginalis to human semen is of short duration, any effect that this fluid may have on the urogenital protozoon could affect its transmission, especially if only few trichomonads are present. Small numbers of parasites (about 2500/ml semen) incubated in semen from different donors at 37 degrees C, were found to survive or grow for up to 12 hours in all samples and for up to 24 hours in most. Survival and growth of T vaginalis in semen most resembled that found in Diamond's trypticase, yeast extract, and maltose (TYM) medium without serum supplement, rather than in complete TYM medium and phosphate buffered saline. Contrary to previous reports, semen did not inhibit the survival of T vaginalis, and the presence of trichomonads did not alter motility or numbers of spermatozoa up to 24 hours. The data suggest that semen provides a favourable milieu for transmitting trichomonads.

Effect of human semen on herpes-simplex virus-2.

This study was designed to evaluate the suggestion of inhibition of infectivity of HSV-2 by seminal fluid by examining relationships between HSV-2 and components of human semen. Viral infectivity was quantitated by TCID50 measurements of CPE in comparing viral inoculated aliquots of whole semen, seminal fluid, and control (EMEM) medium during incubation at 0, 5, and 24 hours. Statistical analysis of data from 15 experiments revealed inhibition by seminal fluid during 5 hours of incubation. After 24 hours viral infectivity was maintained significantly higher with whole semen than with other treatments, suggesting favorable spermatozoal interaction. Electron microscopy revealed no intimate ultrastructural relationship between the virion and the spermatozoon.

Cryosurvival of herpes simplex virus-2 during cryopreservation of human spermatozoa.

Herpes simplex virus (HSV) is a sexually transmitted agent which has potential association with cervical cancer, as well as with high morbidity and mortality in perinatal infection. Its incidence is estimated just after gonorrhea and Chlamydia trachomatis infections. Cryobanking of human semen is accepted worldwide in therapeutic use for both husband and donor. Serious consequences could follow the clinical applications of cryobanked human semen in insemination, if some ejaculates contain HSV and the virus survives the processing of semen in cryopreservation. There is then the likelihood of infection of the female, and through her, the child, generally during parturition.

Effect of isolation by albumin density gradients on ultrastructure of human spermatozoa.

Unlike results with glass wool filtration, based on identical criteria, no evidence was observed in this study for induced ultrastructural damage to the cell membrane and acrosome of heads of spermatozoa during isolation by the albumin density gradient technique. To the contrary, significant numbers of ultrastructurally abnormal spermatozoa were screened out of populations, as were spermatozoa with obvious light-microscopic abnormalities. There was no correlation, however, between structural abnormalities noted with light or electron microscopy and spermatozoal swimming ability. Reduced fertility with inseminated isolated spermatozoa most likely would be due to the tremendous loss (91%) of motile cells rather than cellular injury by the albumin gradient procedure.

Effect of glass wool filtration on ultrastructure of human spermatozoa.

The results of this study strongly suggest that filtration by glass wool can induce damage to the membrane and acrosome of the heads of some spermatozoa in a population. It is possible that the potential fertilizing capacity of a population of human spermatozoa may be reduced as a consequence of these alterations, especially those to the acrosome. The results suggest that sufficient clinical applications of glass wool filtration in artificial insemination is the only way to evaluate both the potential benefits of this process and the potential drawbacks to efficiency that may be caused by a degree of ultrastructural damage.

Importance of frozen-stored human semen in the spread of gonorrhea.

The sensitivity of Neisseria gonorrhea in both infected human semen and urine to routine handling and storage above and below the freezing temperature has been studied. It was found that gonococci in such physiologic media remain viable in storage under atmospheric conditions, in gauze-plugged shell vials or in sealed plastic straws, for 1 to 8 days at +22 degrees C. Furthermore, cryo-survival of gonococci was demonstrated after pretreatment at +22 degrees C; freezing to, and thawing from, -196 degrees C; and frozen storage at -196 degrees C for at least 18 months, with and without the cryo-protectant glycerol. The relationship between laboratory diagnostic testing, disease transmission by males, and therapeutic donor insemination (AID) was discussed. It was strongly urged that only frozen-stored semen should be used in AID, primarily because of the possible use of asymptomatic donors. Cryo-preservation would ensure adequate testing of each sample to be inseminated, preventing the transmission of pandemic gonorrhea via semen.

Synopsis of the use of frozen human semen since 1964: state of the art of human semen banking.

PIP: The history of frozen human semen is reviewed and the clinical devel opments which began in 1964 are given in greater detail. Artificial insemination has become widely accepted and thousands have been performed successfully. 40-45% survival of spermatozoa have been recorded after 10 years of storage. No evidence suggests that storage produces genetic changes. Conservative data show 564 children have been born from frozen semen using approved insemination procedures, 7 abnormal children were born, 50 spontaneous abortions were recorded, and 65 pregnancies had not yet resolved themselves. Periods of preservation ranged from 13 months (Japanese series) to 10 1/4 years (U.S.). Names and addresses of researchers maintaining semen banks are given. Application of the technology to infertility, as "insurance" for men undergoing vasectomy, and as a method for improving genetic characteristics are discussed. Commercial banks with regulated standards and a clarified policy of no guarantee of fertility were endorsed as essential to large-scale application of this technology.^ieng