Fertility Preservation Options Research Articles

Testicular Tissue Vitrification: a Promising Strategy for Male Fertility Preservation.

Destruction of spermatogonial stem cells in juvenile men survivors of pediatric cancers leads to infertility as a side effect of gonadotoxic therapies. Sperm freezing before cancer treatment is commonly used in the clinic for fertility preservation, but this method is not applicable for prepubertal boys due to the lack of mature sperm. In these cases, cryopreservation of testicular tissues is the only option for fertility preservation. Although controlled slow freezing (CSF) is the most common procedure for testicular tissue cryopreservation, vitrification can be used as an alternative method. Controlled vitrification has prevented cell damage and formation of ice crystals. Procedures were done easily and quickly with a brief exposure time to high concentration of cryoprotectants without expensive equipment. Different studies used vitrification of testicular tissues and they assessed the morphology of seminiferous tubules, apoptosis, and viability of spermatogonial cells. Transplantation of vitrified testicular tissue into infertile recipient mice as well as in vitro culture of vitrified tissues was done in previous studies and their findings showed complete spermatogenesis and production of mature sperm. Review articles usually have compared controlled slow freezing with vitrification. In this review, we focused only on the vitrification method and its results. Despite promising results, many studies have been done for finding an optimal cryopreservation protocol in order to successfully preserve fertility in prepubertal boys.

Damages and stress responses in sperm cells and other germplasms during dehydration and storage at nonfreezing temperatures for fertility preservation.

Long-term preservation of sperm, oocytes, and gonadal tissues at ambient temperatures has the potential to lower the costs and simplify biobanking in human reproductive medicine, as well as for the management of animal populations. Over the past decades, different dehydration protocols and long-term storage solutions at nonfreezing temperatures have been explored, mainly for mammalian sperm cells. Oocytes and gonadal tissues are more challenging to dehydrate so little to no progress have been made. Currently, the detrimental effects of the drying process itself are better characterized than the impact of long-term storage at nonfreezing temperatures. While structural and functional properties of germ cells can be preserved after dehydration, a long list of damages and stresses in nuclei, organelles, and cytoplasmic membranes have been reported and sometimes mitigated. Characterizing those damages and better understanding the response of germ cells and tissues to the stress of dehydration is fundamental. It will contribute to the development of optimal protocols while proving the safety of alternativestorage options for fertility preservation. The objective of this review is to (1) document the types of damages and stress responses, as well as their mitigation in cells dried with different techniques, and (2) propose new research directions.

Fertility preservation in the pediatric population-experience from a German Cryobank for ovarian tissue.

Counseling children on the possibility of fertility preservation prior to a gonadotoxic treatment supports the decision-making process, taking into account that the patients are in a very vulnerable and mentally exhausting situation following the diagnosis. Referral to specialists can be optimized on-site by routing slips with contact addresses, phone numbers, and mail contacts; available time slots for consultation; possibly offers for cost coverage; and an easy-to-understand information leaflet about the different options available. Some of the options for fertility preservation in the prepubertal population especially are still experimental. The unique possibility of fertility preservation before the onset of the gonadotoxic therapy, which may cause premature ovarian insufficiency or azoospermia in the future, should be highlighted.

7884 Laparoscopic Ovarian Transposition for Fertility Preservation

<h3>Study Objective</h3> To review the role of ovarian transposition in fertility preservation technologies, describe laparoscopic ovarian transposition performed for fertility preservation in a patient undergoing pelvic radiation, and to discuss the feasibility of this procedure for the general obstetrician gynecologist for cancer patients desiring fertility preservation. <h3>Design</h3> We present a case report of a patient who undergoes laparoscopic ovarian transposition performed for fertility preservation prior to planned pelvic radiation therapy for melanoma metastatic to the sacrum. <h3>Setting</h3> Ambulatory operating room setting with patient in dorsal lithotomy position. <h3>Patients or Participants</h3> Case report of a patient selected from our institution. <h3>Interventions</h3> Laparoscopic ovarian transposition affixes the ovaries out of the pelvis, preventing their irradiation during planned radiation therapy for cancers involving the pelvis. <h3>Measurements and Main Results</h3> A laparoscopic ovarian transposition performed for fertility preservation was successfully performed prior to planned pelvic radiation therapy. <h3>Conclusion</h3> Women undergoing planned pelvic radiation therapy should be offered ovarian transposition prior to starting therapy. Laparoscopic ovarian transposition is an option for fertility preservation that is within the scope of the general obstetrician gynecologist.

The experience of female oncological patients and fertility preservation: A phenomenology study.

The aim of this study is to analyse the experience of fertility preservation among female oncological patients. A phenomenological study was conducted in an Assisted Human Reproduction Unit. The sample was composed of 14 females of reproductive age diagnosed with a cancer that may affect reproductive function, who had undergone a fertility preservation procedure. In-depth interviews were carried out, and the transcripts were entred into the Atlas.ti.v.8.2.3 qualitative analysis software package. The steps proposed by Taylor-Bogdan were followed in the analysis of the data. Four thematic categories emerged in relation to the experience with the fertility preservation process: interruption of life plans and changes in the person; knowledge about fertility preservation options, reproductive capacity and impact because of the possible loss; dyadic relationshipsand social representations of fertility. Patients suffer from difficulties when making decisions about fertility whilst dealing with a cancer diagnosis. They need adequate information and support from health professionals. Despite increasing awareness of fertility preservation, there is a lack of knowledge regarding patients' experiences and needs related to this process.

7578 Hysterectomy with or without Oophorectomy for Transgender Male Patients: Preoperative and Intraoperative Considerations

<h3>Study Objective</h3> To review the preoperative and intraoperative considerations for gynecologic surgeons when performing hysterectomy with or without oophorectomy for transgender male patients. <h3>Design</h3> Stepwise demonstration of techniques with narrated video footage. <h3>Setting</h3> Approximately 0.3% of hysterectomies performed annually in the U.S. are for transgender male patients, most commonly to alleviate gender dysphoria. A hysterectomy with or without oophorectomy can be offered to patients who have met criteria outlined by the World Professional Association for Transgender Health (WPATH) guidelines. <h3>Patients or Participants</h3> Transgender male patients seeking hysterectomy with or without oophorectomy. <h3>Interventions</h3> Important perioperative counseling points for transgender male patients include: •Establishing terminology to minimize gender dysphoria •Discussing options for fertility preservation •Discussing the importance of continuing testosterone therapy post-oophorectomy to reduce the loss of bone density •Reviewing intraoperative and postoperative expectations Important intraoperative considerations for the performance of safe and effective hysterectomy with or without oophorectomy for transgender male patients include: •Maintaining a margin of at least 2 cm between the surgical instrument and the ovary during coagulation and transection of the infundibulopelvic ligament to prevent ovarian remnant syndrome •Performance of a two-layer vaginal cuff closure to reduce the risk of vaginal cuff complications <h3>Measurements and Main Results</h3> The patients recovered well without any postoperative complications. <h3>Conclusion</h3> In conclusion, it is essential to minimize the patient's experience of gender dysphoria by providing gender-affirming care before, during and after surgery. In particular, the surgeon should take care to perform oophorectomy with an adequate margin to minimize the risk of ovarian remnant syndrome and use a two-layer laparoscopic vaginal cuff closure to decrease the risk of vaginal cuff complications.

RF02 | PSAT119 AMH and Spontaneous Puberty in a Diverse US Turner Syndrome Clinic Cohort: A Cross-sectional study

Abstract Background Turner syndrome (TS) is caused by the absence of whole/part of the second X-chromosome. Primary ovarian insufficiency (POI) is common in TS and serum anti-Mullerian hormone (AMH) of &amp;lt;4 pmol/L (0.56 ng/mL) is suggestive of POI and lack of spontaneous puberty. Objective To describe the distribution of AMH and spontaneous puberty in a cross-sectional cohort of patients in a newly-established pediatric TS specialty clinic. Methods Data on all TS patients seen in the multidisciplinary clinic between 1/1/2019 and 10/27/2021 were abstracted from retrospective chart review into a REDCap database. Descriptive analysis, one way ANOVA and two sample t-tests for pair-wise comparisons was completed (SPSSv28.0 and Excelv2102), with statistical significance set at p&amp;lt;0.05. Results Ninety-six patients (9% Asian, 13.5% African American and 37.5% Caucasian, and rest unknown/other self-reported race, and 29.2% Hispanic ethnicity) were enrolled in our clinic. While the majority were diagnosed postnatally (74%, median age 5 years), 6% were prenatal and 20% had positive prenatal screen with postnatal confirmation. Karyotype distribution: 41% non-mosaic monosomy X, 17% Xq (including 14% isochromosome Xq) and 14% Xp structural abnormalities, 10% ringX, 7% 45,X/46,XX mosaicism and 9% with Y-chromosome. Median age at first TS clinic visit was 10 years (IQR 9.7) with median height at 2%ile, weight 31%ile and BMI 73%ile. AMH was measured in 88 patients and only 33 (37.5%) had detectable (&amp;gt;0.015 ng/mL) AMH; 8 (9%) had AMH &amp;gt;0.56ng/mL. Mean AMH was significantly lower in 45,×monosomy (0.03+/- 0.14) vs. other karyotypes (0.44+/-1.13)(p=0.01). A one-way ANOVA revealed a statistically significant difference in AMH between 45,X, structural abnormalities in Xp/Xq and, mosaic 45,X/46,XX karyotypes (p&amp;lt;0.001). Pair-wise comparisons confirmed mean AMH in 45,X/46,XX was significantly higher (3.1ng/mL) (p&amp;lt;0.001) compared with 45,×(0.03ng/mL), Xq (0.02ng/mL) and Xp (0.08ng/mL) abnormalities but no other comparisons were statistically significant. Spontaneous thelarche was reported in 23/58 (39.6%) pubertal patients with mean AMH (0.65ng/mL +/-1.3) compared to absent spontaneous thelarche (N=35, 0.02 +/- 0.04) (p=0.03). Spontaneous menarche was reported in 9 patients, with mean AMH 1.8+/- 1.7ng/mL; 4/9 had 45,X/46, XX karyotype and 1 each had 45,X, Xp, Xq, isoXq, and ringX karyotypes. The mean age at spontaneous thelarche was lower than age at thelarche with hormonal induction (11 + 1 years vs. 14 + 2 years, p &amp;lt; 0.001). Median age at estrogen initiation was 12 years (IQR 3); 86% received transdermal estradiol with menarche at a median age of 15 years. While 78/96 (81%) received fertility preservation counseling at a mean age of 11 + 6 years, only 4 utilized fertility preservation options (1 oocyte cryopreservation and 3 ovarian tissue cryopreservation). Although cross-sectional data limit inferences on AMH as a marker of puberty, our data represents a large and diverse cohort of TS patients in the US. Presentation: Saturday, June 11, 2022 1:00 p.m. - 3:00 p.m., Saturday, June 11, 2022 1:24 p.m. - 1:29 p.m., Saturday, June 11, 2022 1:24 p.m. - 1:29 p.m.

Australian fertility preservation guidelines for people with cancer 2022: review and recommendations.

Australian fertility preservation guidelines for people with cancer 2022: review and recommendations.

Fertility preservation for pediatric patients with hemoglobinopathies: Multidisciplinary counseling needed to optimize outcomes.

Hemoglobinopathies are autosomal recessive disorders that occur when genetic mutations negatively impact the function of hemoglobin. Common hemoglobinopathies that are clinically significant include sickle cell disease, alpha thalassemia, and beta thalassemia. Advancements in disease-modifying and curative treatments for the common hemoglobinopathies over the past thirty years have led to improvements in patient quality of life and longevity for those who are affected. However, the diseases, their treatments and cures pose infertility risks, making fertility preservation counseling and treatment an important part of the contemporary comprehensive patient care. Sickle cell disease negatively impacts both male and female infertility, primarily by testicular failure and decreased ovarian reserve, respectively. Fertility in both males and females with beta thalassemia major are negatively impacted by iron deposition due to chronic blood transfusions. Hematopoietic stem cell transplant (HSCT) is currently the only curative treatment for SCD and transfusion dependent beta thalassemia. Many of the conditioning regimens for HSCT contain chemotherapeutic agents with known gonadotoxicity and whole-body radiation. Although most clinical studies on toxicity and impact of HSCT on long-term health do not evaluate fertility, gonadal failure is common. Male fertility preservation modalities that exist prior to gonadotoxic treatment include sperm banking for pubertal males and testicular cryopreservation for pre-pubertal boys. For female patients, fertility preservation options include oocyte cryopreservation and ovarian tissue cryopreservation. Oocyte cryopreservation requires controlled ovarian hyperstimulation (COH) with ten to fourteen days of intensive monitoring and medication administration. This is feasible once the patient has undergone menarche. Follicular growth is monitored via transvaginal or transabdominal ultrasound, and hormone levels are monitored through frequent blood work. Oocytes are then harvested via a minimally invasive approach under anesthesia. Complications of COH are more common in patients with hemoglobinopathies. Ovarian hyperstimulation syndrome creates a greater risk to patients with underlying vascular, pulmonary, and renal injury, as they may be less able to tolerate fluids shifts. Thus, it is critical to monitor patients undergoing COH closely with close collaboration between the hematology team and the reproductive endocrinology team. Counseling patients and families about future fertility must take into consideration the patient's disease, treatment history, and planned treatment, acknowledging current knowledge gaps.

Autophagy phenomenon in mice ovaries following transplantation

Ovarian cryopreservation and transplantation is a fertility preservation option, but a majority of follicles were lost due to grafting process. Accelerated activation of primordial follicle and follicle apoptosis are responsible for follicle depletion following grafting. Since apoptosis and autophagy share common upstream signals, and cell switches between apoptosis and autophagy, the aim of this study was to investigate whether autophagy took place in ovaries following transplantation. Transmission electron microscopy and immunofluorescence were used to detect autophagic changes in grafted mice ovaries. Results shown that autophagosomes increased rapidly at 2 days following ovarian transplantation in granulosa cells of growing follicles, and stromal cells. The study indicated that autophagy is actively involved during ovarian grafting. Manipulation of autophagy might provide another way to improve ovarian reserved during grafting.

A CELL THERAPY ASSISTED NOVEL MICROFLUIDIC DEVICE PROMOTESIN VITROSPERMATOGENESIS IN NEONATAL MICE

Chemo-radiotherapy applications result in permanent infertility in half of male pediatric cancer survivors1. Spermatogonial stem cells (SSC) constitute the only option for fertility preservation since spermatogenesis is not initiated yet2. The rationale of study is that bone marrow derived mesenchymal stem cells (BMMSC) have similar embryonic origin and gene expression profile with Sertoli cells3,4 that play crucial role in proliferation of SSCs, and microfluidic devices (MD) simulate microvascular flow in the body that could promote for in vitro spermatogenesis (IVS). We hypothesized that, syngeneic BMMSC-conditioned medium (BMMSC-CM) contributed monolayer pumpless MD leads to survival, expansion and differentiation of SSC pool in vitro.

DELAYING CHILDBEARING TO BECOME A DOCTOR: AN EXPLORATION OF MEDICAL STUDENT FAMILY PLANNING GOALS AND KNOWLEDGE OF FERTILITY PRESERVATION OPTIONS

Women pursuing medical education delay child-bearing for many reasons, with career being one of the most salient reasons. Female fecundity declines with age, but the average age of women matriculants to medical school continues to increase. Our study investigated the reproductive goals of medical students and their knowledge and attitudes regarding fertility preservation options.

Fertility preservation: knowledge, awareness, and attitude of university students and professors.

Delayed pregnancy is a worldwide trend, especially in Western countries. University students and professors are at high risk of presenting age-related reproductive difficulties due to this new reproductive profile. Thus, through this study, we aimed at exploring the knowledge, awareness, and attitude of university students and professors related to fertility and fertility preservation (FP). We adopted a prospective cross-sectional study design and included students and professors from private university located in the Northeast of Brazil. Eligible participants (male and female) were invited through an online message. The participants accessed the online questionnaire through a link. We performed 256 surveys (100 students and 156 professors). The overall mean age of participants was 35.8±13.1 years (from 18 to 67 years). Fertility was considered relevant by all participants, being very important among a greater number of students compared to professors, 61% versus 30.1%, P<0.001, respectively. The main reasons why participants could have postponed parenthood were reach financial stability (62.1%), career building (51.2%), health issues (37.9%), and not having a partner (33.9%). Students demonstrated a better understanding of FP and highlighted the importance of the age of females at the time of the oocyte cryopreservation. Very few students and professors already discussed reproductive planning with a health professional. We observed a deficiency in the knowledge of Brazilian university students and professors about female fertility and FP options. Thus, exposing the population to information related to FP should be hyped in the university environment.

TURNERFERTILITY STUDY: FERTILITY PRESERVATION IN YOUNG GIRLS WITH TURNER SYNDROME BY FREEZING OVARIAN CORTEX TISSUE

Turner syndrome (TS) is a heterogeneous chromosomal disorder affecting one in 2,500 live-born girls. Most girls with TS are confronted with infertility at an early age. Over the past decades, ovarian tissue cryopreservation (OTC) has been successfully performed and is well documented for young females facing fertility-threatening anti-cancer treatments. In this group, OTC is now a valid fertility preservation (FP) option. OTC might also be a promising technique for FP in girls with TS, as it can be performed regardless of the patient’s age.

INFLUENCE OF FERTILITY PRESERVATION INDICATION ON OOCYTE AND EMBRYO CRYOPRESERVATION: A COMPARISON OF MEDICALLY INDICATED VS ELECTIVE CYCLES

Cryopreservation of embryos/oocytes are feasible fertility preservation (FP) options in patients facing gonadotoxic treatments and age-related fertility decline. A paucity of data exists to guide patient counseling about cycle outcomes particularly ones pursing FP for medical indications. The objective of this study is to compare expected oocyte/embryo yield and cycles between medically indicated and elective FP groups.

FERTILITY PRESERVATION TECHNIQUES WITH OR WITHOUT OVARIAN STIMULATION DO NOT CHANGE DISEASE-FREE SURVIVAL OF YOUNG BREAST CANCER PATIENTS

Fertility is often impaired in young women treated for breast cancer (BC). Therefore, fertility preservation (FP) has become a major issue in this population. Cryopreservation of oocytes or embryos after controlled ovarian hyperstimulation (COH) represents the most established method in this clinical situation. However, the hormonal consequences of COH protocols still raise safety concerns, often leading oncologists to contraindicate the use of this FP technique. Although alternative FP options without exogenous hormone administration may be considered, they remain suboptimal for treating the putative future infertility.

Fertility Preservation and Reproductive Potential in Transgender and Gender Fluid Population

The gender diverse and transgender community is a minor patient group that is encountered with increasing frequency in the clinical setting, attributed to the improved awareness and access to medical facilities. Partial impairment to permanent elimination of fertility potential and outcomes depending on the treatment modality usually is a result of gender-affirming therapy, which includes both hormone therapy and surgical intervention. Although seldom conducted in the clinical field, transgender patients should be counseled on their fertility preservation options prior to medical and surgical gender transition. There is relatively limited data and clinical information regarding fertility preservation for transgender individuals. Current treatment regimens are based on protocols from fertility preservation after oncological treatments. Major barriers for the transgender population exist due to the lack of information provided and clinical narrative that is not familiar to the physician or health care provider, although there are various options for fertility preservation. A deeper understanding of this clinical agenda and the mandatory processes will ultimately result in a much more comprehensive and specific care for transgender individuals who are in great need for fertility counseling or treatment options that concern fertility preservation. In this review, current clinical approaches will be summarized and fertility preservation options along with ongoing and future clinical trials in fertility preservation for transgender individuals will be thoroughly reviewed.

Testicular tissue cryopreservation for fertility preservation in prepubertal and adolescent boys: A 6 year experience from a Swiss multi-center network.

Testicular tissue cryopreservation is the only option of fertility preservation in prepubertal boys. While it is considered experimental, since procedures to obtain mature spermatozoa from prepubertal testicular tissue are still under development, testicular tissue cryopreservation programs have emerged worldwide. Our aim was to study the feasibility and safety of a program of testicular tissue cryopreservation in prepubertal and adolescent boys facing gonadotoxic treatment in three University hospitals in Switzerland. Testicular tissue cryopreservation was accepted by 90% of families, with a total of 35 patients included. The average patient age was 8.5 years (range 7 months to 18.5 years). Malignancies were the most common diagnosis (31 patients, 88.6%) with 16 (45.7%) solid tumors and 15 (42.9%) hematological malignancies. Four (11.4%) patients had a benign condition. The main indication for testicular tissue cryopreservation was conditioning for hematologic stem cell transplantation (25 patients, 71.4%). Testicular tissue was cryopreserved according to the freezing protocol of Louvain Catholic University (Belgium), which includes either only immature testicular tissue freezing, or mature and immature testicular tissue freezing depending on the age of the patient and the presence or absence of haploid cells. The median number of spermatogonia per tubule cross-section was 2 (range 0–6) and spermatozoa were found in only one patient. Tumoral cells were found in one testicular biopsy of a leukemic patient. There were two minor adverse events and none of them required medical treatment or surgical revision. Five patients died during follow-up. Our data demonstrate the feasibility and safety of a program of testicular tissue cryopreservation coordinated by a multidisciplinary team of fertility preservation. Despite the experimental aspect of the procedure, the acceptation rate was high, which highlights the willingness of families and patients to participate in testicular tissue cryopreservation.

NMN Supplementation Rescues Fertility and Bone Strength in Chemotherapy-Treated Mice

Background: Cancer survivors often face infertility as a result of ovarian toxicity from cytotoxic chemotherapy drugs, which trigger a cascade of events ultimately causing follicular reserve exhaustion and endocrine disruption. Apart from infertility, this can have severe consequences on women’s health with early-onset menopause and a decline in bone health. For girls with cancer, ovarian tissue cryopreservation (OTC) is the only option for fertility preservation. OTC + transplantation may provide future fertility potential, but it provides minimal/no protection from endocrine failure and osteoporosis. Aim: To assess the effects of the nicotinamide adenine dinucleotide (NAD+) precursor nicotinamide mononucleotide (NMN) on ovarian function, fertility, and late-life bone health. Method: 7-day old female mice were treated +/- cisplatin (2mg/kg). Two weeks later, +/- NMN was delivered in drinking water (2g/L) and sustained throughout life. A breeding trial was conducted at 6 weeks, and to assess late-life effects on bone structure, mice were maintained until euthanasia at 24 months of age. Results: Cisplatin caused a dramatic decline in all fertility endpoints. NMN supplementation rescued breeding performance in cisplatin treated animals by 5-fold (p=0.015), as measured by the cumulative number of pups/female. Given the importance of ovarian function to estrogen release and the risk of osteoporosis, we sought to determine the impacts of these interventions on late-life bone health. Bones were subject to mechanical and structural analysis to assess differences in the onset of osteoporosis. In cisplatin treated animals, NMN rescued cortical bone thickness (femur diaphysis p[Formula: see text]0.0001, femur metaphysis p=0.0081, tibia p=0.0072), bone volume (femur p=0.0015, tibia p=0.0102), and bone density (femur p=0.0463) to control levels, and increased mechanical strength (tibia, p=0.0024). Conclusion: Long-term NMN treatment delivered following chemotherapy protected against subsequent ovarian failure and infertility, and notably improved late-life bone health, possibly due to the prevention of premature ovarian failure.

Post-Thaw Sperm Recovery Rates; A Retrospective Study

Background: Cryopreservation is currently the only fertility preservation option available to patients undergoing gonadotoxic treatment. Much research in the field focusses on techniques, media and equipment employed to cryopreserve cells, and demonstrating cell viability after using such. Comparatively, there is a death of information available on the procedures and equipment used to thaw cryopreserved cells, or estimates for post-thaw recovery. Aim: This is a retrospective study assessing semen parameters in post-thawed specimen from patients who attended the Royal Women’s Hospital Andrology Unit for sperm cryopreservation prior to gonadotoxic or other treatments. The study aims to examine the relationship between sperm cryopreservation technique, media and period of storage with post-thaw sperm parameters. Method: Patients with written consent to discard their stored semen were selected for inclusion in the study (n=59); samples with sperm concentration [Formula: see text]1 million/ml prior to freezing (n=11) were excluded. Semen samples were thawed accordingly: 5 minutes at room temperature followed by incubation at 37[Formula: see text]C for sperm straws; vials were transferred directly to a mixer incubator at 37[Formula: see text]C. All samples were immediately assessed for sperm concentration and motility. Recovery rates were calculated and data were analysed using SPSS (IBM, USA) version 25. Results: Progressive motility recovery was 32.4% ± 20.6. Patients were divided into groups according to period of storage, and showed no significant differences for recovery of motility and concentration (p=0.44, 0.96, respectively). Furthermore, progressive motility recovery rates were not significantly different between freezing techniques (p=0.116) and media (p=0.099). Conclusion: Data from this study are preliminary and further study is warranted to fully extrapolate the relationship between sperm cryopreservation protocols and period of storage on recovery rates. The lack of relationship between these factors exhibited in the data could suggest that research and development in thaw technologies may be more pertinent to improving sperm recovery rates.