Modified permeable cryoprotectant-free vitrification method for three or fewer ejaculated spermatozoa from cryptozoospermic men and 7-year follow-up study of 14 children born from this method.

STUDY QUESTION: What technique can be used to successfully cryopreserve three or fewer ejaculated spermatozoa from cryptozoospermic men and is the physical and cognitive development of children born after this technique normal? SUMMARY ANSWER: The modified cryopreservation method for three or fewer human spermatozoa from cryptozoospermic men showed a recovery rate above 95% and a survival rate just under 90%, and the physical and cognitive abilities of the children born after ICSI were comparable to those born after natural conception. WHAT IS KNOWN ALREADY: Clinical outcomes of ICSI using cryptozoospermic men's ejaculated spermatozoa are considered to be inferior to that using testicular spermatozoa from microsurgical testicular sperm extraction (Micro-TESE), possibly because the DNA fragmentation rate is higher in ejaculated spermatozoa than in testicular spermatozoa from Micro-TESE. STUDY DESIGN, SIZE, DURATION: Evaluation of the efficiency of cryopreservation of three or fewer spermatozoa was conducted retrospectively at St. Mother Clinic. The physical and cognitive development of children born after this method was studied between 2011 and 2018. PARTICIPANTS/MATERIALS, SETTING, METHODS: This study included 28 cryptozoospermic men who had three or fewer morphologically normal and motile spermatozoa in their ejaculate after centrifugation and who preferred using cryopreserved spermatozoa to Micro-TESE. Control subjects were 31 cryptozoospermic patients using fresh spermatozoa from their ejaculates and 20 non-obstructive azoospermic patients with fewer than 10 spermatozoa obtained by TESE and vitrified. Clinical outcomes among three groups, vitrified spermatozoa from the ejaculate, fresh spermatozoa from the ejaculate and vitrified spermatozoa from the testis, were statistically analysed. For the 7-year follow up study of the 14 children born after ICSI using the ejaculated vitrified spermatozoa, the Japanese government-issued Boshi Kenko Techo (Mother-Child Handbook) and Kinder Infant Development Scale (KIDS scale) were used to determine whether their physical and cognitive development was comparable to that of naturally conceived children. MAIN RESULTS AND THE ROLE OF CHANCE: Recovery and survival rates were 97.8% (510/521) and 87.1% (444/510) for vitrified spermatozoa from the ejaculate and 92.7% (152/164) and 60.5% (92/152) for vitrified spermatozoa from the testis. Clinical pregnancies (%), miscarriages (%) and live birth rates (%), respectively, among the three groups were as follows: vitrified spermatozoa from the ejaculate: 15(25.0), 2(13.3), 13(21.7); fresh spermatozoa from the ejaculate: 26(24.3), 5(19.2), 20(18.7); and vitrified spermatozoa from the testis: 3(16.7), 0(0.0), 3(16.7). Among the groups, there were no statistically significant differences except for the sperm survival rate and the oocyte fertilisation rate, which were lower for vitrified spermatozoa from the testis compared with vitrified spermatozoa from the ejaculate. The 7-year follow-up study showed that the physical and cognitive development of 14 children born after ICSI using vitrified ejaculated spermatozoa from the ejaculate was comparable to that of naturally conceived children. LIMITATIONS, REASONS FOR CAUTIONS: The maximum number of spermatozoa to which this method can be applied successfully is about 10. When the number of aspirated spermatozoa is over 10, some of them change direction after colliding with each other inside the aspiration pipette and reach the mineral oil, and once this happens, they cannot be expelled out of the pipette. Even though we did not find evidence of DNA fragmentation, further studies with larger participant numbers and longer time periods are necessary. WIDER IMPLICATIONS OF THE FINDINGS: This technique is very useful for the cryopreservation of very small numbers of testicular spermatozoa (fewer than 10) in order to avoid or reduce Micro-TESE interventions. STUDY FUNDING/COMPETING INTEREST(S): No external funding was received to undertake this study. There are no competing interests. TRIAL REGISTRATION NUMBER: N/A.

Ninety babies born after round spermatid injection into oocytes: survey of their development from fertilization to 2 years of age.

OBJECTIVE: To compare physical and cognitive development of babies born after round spermatid injection (ROSI) with those born after natural conception. DESIGN: Comparison of efficiencies of ROSI and ICSI using testicular spermatozoa, performed in the St. Mother Clinic. Physical and cognitive development of ROSI babies recorded by parents in the government-issued Mother-Child Handbook was checked and verified by attending pediatricians. Data included baby's weight gain and response to parents' voice/gesture. SETTING: Assisted reproduction technology practice. PATIENT(S): A total of 721 men participated in ROSI; 90 ROSI babies were followed for 2 years for their physical and cognitive development. Control subjects were 1,818 naturally born babies. INTERVENTION(S): Surgical retrieval of spermatogenic cells from testes; selection and injection of round spermatids into oocytes; oocyte activation, in vitro culture of fertilized eggs, and embryo transfer to mothers. MAIN OUTCOME MEASURE(S): Physical and cognitive development of ROSI babies (e.g., body weight increase, response to parents, and understanding and speaking simple language) compared with naturally born babies. RESULT(S): Of 90 ROSI babies, three had congenital aberrations at birth, which corrected spontaneously (ventricular septa) or after surgery (cleft lip and omphalocele). Physical and cognitive development of ROSI babies was similar to those of naturally born babies. CONCLUSION(S): There were no significant differences between ROSI and naturally conceived babies in either physical or cognitive development during the first 2 years after birth. CLINICAL TRIAL REGISTRATION NUMBER: UMIN Clinical Trials Registry UMIN000006117.

Fourteen babies born after round spermatid injection into human oocytes.

During the human in vitro fertilization procedure in the assisted reproductive technology, intracytoplasmic sperm injection is routinely used to inject a spermatozoon or a less mature elongating spermatid into the oocyte. In some infertile men, round spermatids (haploid male germ cells that have completed meiosis) are the most mature cells visible during testicular biopsy. The microsurgical injection of a round spermatid into an oocyte as a substitute is commonly referred to as round spermatid injection (ROSI). Currently, human ROSI is considered a very inefficient procedure and of no clinical value. Herein, we report the birth and development of 14 children born to 12 women following ROSI of 734 oocytes previously activated by an electric current. The round spermatids came from men who had been diagnosed as not having spermatozoa or elongated spermatids by andrologists at other hospitals after a first Micro-TESE. A key to our success was our ability to identify round spermatids accurately before oocyte injection. As of today, all children born after ROSI in our clinic are without any unusual physical, mental, or epigenetic problems. Thus, for men whose germ cells are unable to develop beyond the round spermatid stage, ROSI can, as a last resort, enable them to have their own genetic offspring.

Recurrent Male Breast Cancer Accompanied by Carcinomatous Pleuritis That Responded to Combination Therapy with High-Dose Toremifene and Docetaxel.

BACKGROUND: Male breast cancer (MBC) is a rare disease with no standard treatment compared to female breast cancer. There are very few reports that go beyond second-line chemotherapy and endocrine therapy for advanced and recurrent MBC. CASE REPORT: This report presents a case of recurrent MBC accompanied by carcinomatous pleuritis that responded to combination therapy with high-dose toremifene (TOR) and docetaxel (DOC). A 63-year-old male patient had previously undergone a modified radical mastectomy for left breast cancer and received several series of systemic chemotherapy and endocrine therapy. He complained of severe dyspnea, and was admitted to our hospital due to a massive left pleural effusion caused by carcinomatous pleuritis. He received combination therapy with high-dose TOR and biweekly DOC. The pleural effusion disappeared without any severe side effects after 4 cycles of the therapy. Thereafter, his disease stabilized for 1 year without re-increase of tumor markers under continuous treatment with the combination therapy. CONCLUSIONS: High-dose TOR and DOC might be effective even beyond second-line chemotherapy and endocrine therapy for MBC to overcome potential multiple drug resistance and diminish inevitable side effects.