FERTI.NET HIGHLIGHTS Issue 1 - Week 2 - Volume 4 - 2002 Ferti.Net Highlights Archive This fortnightly news service brings you the latest news on Assisted Reproduction Techniques as they have been reported in the media. Sources include on-line media, medical data bases, original press releases, trade journals, national daily newspapers and broadcasts reports, as well as peer-reviewed publications. It also keeps you up-to-date with the latest issue of the Ferti.Net Magazine. -------------------------------------------------------------------------------- Index a.. The fine structure of human embryonic stem cells b.. Blastocyst culture in human IVF: the final destination or a stop along the way? c.. Determinants of early follicular phase gonadotrophin and estradiol concentrations in women of late reproductive age d.. Genetic risk factors in infertile men with severe oligozoospermia and azoospermia e.. Is your IVF programme good? f.. Restoration of Spermatogenesis and Fertility in Azoospermic Mutant Mice by Suppression and Reelevation of Testosterone Followed by Intracytoplasmic Sperm Injection g.. It's a knockout! Male infertility and neuropathology h.. The role of testicular biopsy in the modern management of male infertility -------------------------------------------------------------------------------- The fine structure of human embryonic stem cells Dr Henry Sathananthan1, Dr Martin Pera2, Alan Trounson3, 1Monash Institute of Reproduction & Development, 27-31 Wright Street, Clayton, Vic. 3168, Australia, 2Monash Institute of Reproduction & Development, Monash University, Melbourne, Australia, 3Monash Institute of Reproduction and Development, Monash University, Melbourne, Australia The fine structure of human embryonic stem (ES) cell colonies were analysed by transmission electron microscopy (TEM) after 35 passages of in-vitro culture. Most cells formed compact, saucer-shaped colonies with epithelioid cells on the periphery and polygonal cells within the colony. Three morphological types of cells were identified based on their fine structure: undifferentiated cells resembling inner cell mass (ICM) cells of blastocysts; protein-synthesizing cells at the onset of cellular differentiation; and compact masses of secretory cells resembling unicellular goblet cells of the intestine. The predominant cell type was the undifferentiated ES cells resembling ICM cells of blastocysts. These cells had large nuclei containing reticulated nucleoli, well-developed rough endoplasmic reticulum (RER), Golgi complexes, elongated tubular mitochondria, lysosomes and typical centrosomes with centrioles associated with microtubules and microfilaments, organizing the cytoskeleton. The isolated or attached protein-synthesizing cells at the onset of differentiation had extensive RER and large Golgi complexes. The morphologically differentiated cells formed compact colonies and resembled goblet-like cells in microstructure. They had RER and large Golgi complexes associated with secretory vesicles. The epithelioid cells at the periphery were columnar and largely polarized by centrosomes associated with Golgi complexes. Epithelioid cells in all three categories had specialized cell junctions (desmosomes), anchored by tonofilaments, and surface blebs. Isolated cells were seen on the surface, towards the centre of the colony, and their free surfaces had microvilli and larger blebs. Approximately 3-5% of all cells were mitotic, with typical bipolar spindles organized by centrosomes, pivotally located at the poles, and appeared to resemble typical somatic cells. Journal Reproductive BioMedicine Online, volume 4, issue 1 Source: www.rbmonline.com Back to Top -------------------------------------------------------------------------------- Blastocyst culture in human IVF: the final destination or a stop along the way? Smith AL. Fertility Lab Consulting, San Antonio, Texas, USA. In the field of human IVF, culturing embryos to the blastocyst stage has gained popularity within the past few years. The impetus to transfer blastocysts has been spurred by several factors: 1) the desire to improve implantation rates in infertility patients, 2) a desire to reduce the multiple pregnancy rate by transferring fewer embryos, 3) the desire to perform pre-implantation genetic diagnosis, and 4) the advent of sequential media. Although culturing human embryos to the Hastocyst stage has improved implantation rates and reduced the incidence of multiple pregnancies in some patient populations, it has not worked for all populations of infertility patients. Factors that may affect the ability of a human embryo to reach the blastocyst stage include the patient's age, cohort of ova retrieved, the use of intracytoplasmic sperm injection of blastomere biopsy, culture conditions, or intrinsic factors within the embryo itself. Culture of human embryos to the blastocyst stage can be an effective method for improving implantation rates and reducing the high order multiple pregnancy rates seen in human IVF clinics when more than three embryos are transferred. Theriogenology 2002 Jan 1;57(1):97-107 Source: www.ncbi.nlm.nih.gov Back to Top -------------------------------------------------------------------------------- Determinants of early follicular phase gonadotrophin and estradiol concentrations in women of late reproductive age. Cramer DW, Barbieri RL, Fraer AR, Harlow BL. Obstetrics and Gynecology Epidemiology Center and Department of Obstetrics and Gynecology, Brigham and Women's Hospital, Boston, MA 02115, USA. BACKGROUND: FSH and estradiol measured during the menstrual (basal) phase of cycles predict the success of infertility treatment; but the role of these hormones as markers for ovarian reserve in normal populations needs further study. METHODS AND RESULTS: From a cohort study of depressed and non-depressed women, a subset of 406 non-depressed women between the ages of 36 and 45 years with spontaneous periods were selected and their concentrations and determinants of basal hormones measured at study entry, 6 and 12 months later were described. FSH and LH increased significantly over the 12 months of observation (P Hum Reprod 2002 Jan;17(1):221-7 Source: humrep.oupjournals.org Back to Top -------------------------------------------------------------------------------- Genetic risk factors in infertile men with severe oligozoospermia and azoospermia. Dohle GR, Halley DJ, Van Hemel JO, van Den Ouwel AM, Pieters MH, Weber RF, Govaerts LC. Andrology Unit, Department of Urology, Department of Clinical Genetics and Department of Obstetrics and Gynaecology, Erasmus University Medical Centre, Rotterdam, The Netherlands. BACKGROUND: Male infertility due to severe oligozoospermia and azoospermia has been associated with a number of genetic risk factors. METHODS: In this study 150 men from couples requesting ICSI were investigated for genetic abnormalities, such as constitutive chromosome abnormalities, microdeletions of the Y chromosome (AZF region) and mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. RESULTS: Genetic analysis identified 16/150 (10.6%) abnormal karyotypes, 8/150 (5.3%) AZFc deletions and 14/150 (9.3%) CFTR gene mutations. An abnormal karyotype was found both in men with oligozoospermia and azoospermia: 9 men had a sex-chromosomal aneuploidy, 6 translocations were identified and one marker chromosome was found. Y chromosomal microdeletions were mainly associated with male infertility, due to testicular insufficiency. All deletions identified comprised the AZFc region, containing the Deleted in Azoospermia (DAZ) gene. CFTR gene mutations were commonly seen in men with congenital absence of the vas deferens, but also in 16% of men with azoospermia without any apparent abnormality of the vas deferens. CONCLUSIONS: A genetic abnormality was identified in 36/150 (24%) men with extreme oligozoospermia and azoospermia. Application of ICSI in these couples can result in offspring with an enhanced risk of unbalanced chromosome complement, male infertility due to the transmission of a Y-chromosomal microdeletion, and cystic fibrosis if both partners are CFTR gene mutation carriers. Genetic testing and counselling is clearly indicated for these couples before ICSI is considered. Hum Reprod 2002 Jan;17(1):13-6 Source: humrep.oupjournals.org Back to Top -------------------------------------------------------------------------------- Is your IVF programme good? Alper MM, Brinsden PR, Fischer R, Wikland M. Boston IVF, Waltham, MA, USA, Bourn Hall Clinic, Cambridge, UK, Fertility Center Hamburg, Hamburg, Germany and Fertility Center Gothenburg, Gothenburg, Sweden. Few standards exist today to assess the quality of an IVF centre. Although much focus is placed upon pregnancy rates, emphasis on this outcome alone is inadequate. The purpose of this report is to examine those factors that should be considered in assessing the overall quality of an IVF centre. Current methods to assess quality are reviewed. Many governing bodies throughout the world currently focus solely on pregnancy rates, which can be misguided if factors such as multiple pregnancies, ovarian hyperstimulation, patient satisfaction, and the proper evaluation of laboratory and clinical protocols are not taken into account. Measurements of quality and methods to improve it are critical in all business models, including IVF. We propose an international standard such as the ISO 9001 for IVF centres to properly evaluate and improve the delivery of their care. Hum Reprod 2002 Jan;17(1):8-10 Source: humrep.oupjournals.org Back to Top -------------------------------------------------------------------------------- Restoration of Spermatogenesis and Fertility in Azoospermic Mutant Mice by Suppression and Reelevation of Testosterone Followed by Intracytoplasmic Sperm Injection. Tohda A, Okuno T, Matsumiya K, Okabe M, Kishikawa H, Dohmae K, Okuyama A, Nishimune Y. Department of Urology, Osaka University Medical School, Suita 565-0871, Japan. Genome Information Research Center, Osaka University, Suita 565-0871, Japan. Department of Food Science and Nutrition, Mukogawa Women's University, Nishinomiya 663-8558, Japan. Department of Science for Laboratory Animal Experimentation, Research Institute for Microbial Diseases, Osaka University, Suita 565-0871, Japan. Advances in assisted reproduction techniques such as in vitro fertilization and intracytoplasmic sperm injection have made paternity possible for many patients with male infertility. However, at least some sperm or spermatids are required for these techniques to be successful, and patients incapable of producing spermatids cannot be helped. Male mice homozygous for the mutant juvenile spermatogonial depletion (jsd) gene show spermatogonial arrest and an elevated intratesticular testosterone level like many other experimental infertility models such as those with iradiation- or chemotherapy-induced testicular damage. In this category of infertile males, suppression of the testosterone level induces spermatogonial differentiation to the stage of spermatocytes but no further. In the present study with jsd mutant mice, we induced spermatogenesis first to spermatocytes and then to elongated spermatids by suppression of testosterone levels with a GnRH antagonist, Nal-Glu, at a dose of 2500 &mgr;g kg(-1) day(-1) for 4 wk and then withdrawal of Nal-Glu. Spermatids were seen in the cross-sections of seminiferous tubules in all mice treated by administration and subsequent withdrawal of Nal-Glu. Four weeks after withdrawal of Nal-Glu, some of the germ cells differentiated into elongated spermatids. Supplementation with testosterone and Nal-Glu after 4 wk of treatment with Nal-Glu alone also induced spermatogenesis similar to the induction by withdrawal of Nal-Glu. Thus, we ascribe the restoration of the differentiation of spermatocytes to spermatids to reelevation of the testosterone level. Furthermore, we successfully rescued male sterility in jsd mice by subsequent intracytoplasmic sperm injection using the elongated spermatids induced by the programmed hormone therapy. Biol Reprod 2002 Jan;66(1):85-90 Source: www.biolreprod.org Back to Top -------------------------------------------------------------------------------- It's a knockout! Male infertility and neuropathology. Anagnostopoulos AV. The Jackson Laboratory, Bar Harbor, ME, USA An increasing number of genetically engineered animals are produced worldwide for use in both basic and applied research. Here, I provide an update of some of the latest mouse knockouts in The Jackson Laboratory Transgenic/Targeted Mutation Database (TBASE), concentrating on those associated with male infertility and neuropathology. Trends Genet 2002 Jan 1;18(1):8-10 Source: journals.bmn.com Back to Top -------------------------------------------------------------------------------- The role of testicular biopsy in the modern management of male infertility. Schoor RA, Elhanbly S, Niederberger CS, Ross LS. Department of Urology, University of Illinois at Chicago, Chicago, IL, USA. PURPOSE: We evaluate the traditional role of isolated testicular biopsy as a diagnostic tool, as opposed to the value as a therapeutic procedure for azoospermic men. MATERIALS AND METHODS: The medical records of azoospermic patients who were evaluated, and treated between 1995 and 2000 were retrospectively analyzed for history, physical examination findings, endocrine profiles, testicular histology and sperm retrieval rates. Based on these parameters, cases were placed into diagnostic categories that included obstructive or nonobstructive azoospermia. Diagnostic parameters used to distinguish obstructive from nonobstructive azoospermia were subjected to statistical analysis with the t-test, analysis of variance and receiver operating characteristics curve. RESULTS: A total of 153 azoospermic men were included in our analysis. Of men with obstructive azoospermia 96% had follicle-stimulating hormone (FSH) 7.6 mIU/ml. or less, or testicular long axis greater than 4.6 cm. Conversely, 89% of men with nonobstructive azoospermia had FSH greater than 7.6 mIU/ml., or testicular long axis 4.6 cm. or less. Receiver operating characteristics analysis revealed that FSH, testicular long axis, and luteinizing hormone were the best individual diagnostic predictors, with areas 0.87, 0.83 and 0.79, respectively. CONCLUSIONS: In the vast majority of patients obstructive azoospermia may be distinguished clinically from nonobstructive azoospermia with a thorough analysis of diagnostic parameters. Based on this result, we believe that the isolated diagnostic testicular biopsy is rarely if ever indicated. Men with FSH 7.6 mIU/ml. or greater, or testicular long axis 4.6 cm. or less may be considered to have nonobstructive azoospermia and counseled accordingly. These men are best treated with therapeutic testicular biopsy and sperm extraction, with processing and cryopreservation for usage in in vitro fertilization and intracytoplasmic sperm injection if they accept advanced reproductive treatment. Diagnostic biopsy is of no other value in this group. Men with FSH 7.6 mIU/ml. or less, or testicular long axis greater than 4.6 cm. may elect to undergo reconstructive surgery with or without testicular biopsy and sperm extraction, or testicular biopsy and sperm extraction alone depending on their reproductive goals. J Urol 2002 Jan;167(1):197-200 Source: www.jurology.com