Assessment of different isolation procedures for blastomeres from two-cell mouse embryos

As an extension of in-vitro fertilization and embryo transfer, detection of genetic and metabolic defects prior to implantation might be possible in the future. The objective of pre-implaiitation diagnosis would be to sample a minimal amount of cellular material of the conceptus for diagnosis prior to transfer. Different protocols for isolating individual blastomeres from two-cell mouse embryos were evaluated. Two-cell mouse embryos were collected and the zona pdlucida was removed by enzyme treatment (pronase), by exposure to acid Tyrode (pH = 2.5) or by mechanical force (suction into a small pipette, removal with a microblade). Individual blastomeres were obtained by exposure to a chelating agent (EDTA-glycme mixture), to Ca 2+-Mg 2+-free PBS or after isolation by mechanical force (bisection with a microblade or suction in a small pipette). The isolated blastomeres were then cultured in vitro without zonae pellucidae. All isolation procedures had a negative impact on the growth patterns of the isolated blastomeres. Different abnormalities could be observed at the blastocyst stage including embryos lacking visible compaction features, embryos with double blastocoeJk cavities and embryos with no inner cell mass (trophoblastic vesicles). © 1987 IRL Press Ltd.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Activation of mouse oocytes, fertilization and development of mouse embryos in vitro after staining with trypan blue or fluorescein diacetate

The influence of vital staining with trypan blue or fluorescein diacetate on the fertilization of mouse oocytes and the developmental potential of mouse embryos was assessed. Neither stain induced spontaneous activation in mouse oocytes, nor did they impair the in vitro development and implantation of mouse zygotes, two-cell embryos, stressed morulae or blastocysts. However, fertilization and subsequent development of mouse oocytes have been shown to be reduced by vital staining. © 1992 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Intracytoplasmic sperm injection

Intracytoplasmic sperm injection (ICSI) with ejaculated, epididymal or testicular spermatozoa was first successful in 1992 and has since become the widely accepted treatment for couples with severe male-factor infertility. The outcome of several thousands of ICSI cycles in terms of fertilization, embryo cleavage and implantation is similar to that for conventional in-vitro fertilization in couples with tubal or idiopathic infertility. To evaluate the important issue of safety of the new technique of ICSI, a prospective follow-up study of children born after ICSI was carried out. The aim was to compile data on karyotypes, congenital malformations, growth parameters and developmental milestones. Parents' agreement to genetic counseling was obtained, as well as prenatal diagnosis, followed by a physical examination of the children at 2 months, 1 and 2 years. Important outcome data to be examined comprise information on major and minor congenital malformations obtained prenatally or after birth, as well as on the further development of the children. © 2002 Elsevier Science Ireland Ltd. All rights reserved.SCOPUS: cp.jinfo:eu-repo/semantics/publishe

Infertilites masculines d'origine genetique

Several etiological factors in male infertility have been identified and it is now clear that a proportion of these factors have a genetic basis. This implies that using assisted reproductive technology procedures and especially intracytoplasmic sperm injection these genetic factors may be transmitted to the children. The following genetic factors involved in severe male infertility will be discussed: the presence of numerical and structural chromosomal anomalies, the occurrence of microdeletions on the long arm of the Y chromosome and the relation between cystic fibrosis and congenital bilateral absence of the vas deferens.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Biochemical and statistical methods

info:eu-repo/semantics/publishe

Effect of 1, 25-dihydroxyvitamin D3 and nifedipine on prolactin release in normal man

In normal man 1, 25 (OH)2-vitamin D3 [1, 25 (OH)2D] increases both basal and TRH-stimulated prolactinemia; this effect is completely reversible by the calcium antagonist nifedipine. Similarly the 1, 25 (OH)2D-induced hypercalcemia is totally inhibited by nifedipine. These findings suggest that both biological effects of 1, 25 (OH)2D are mediated by calcium-dependent mechanisms. © 1985, Italian Society of Endocrinology (SIE). All rights reserved.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Artificial insemination by donor: An alternative for single women

Artificial insemination using donor spermatozoa (AID) has been known as a treatment for heterosexual couples where the husband has been found to be infertile. The first woman without a male partner wanting a child by AID was admitted to the fertility centre in 1981. In the literature, two categories of alternative requests, i.e. those from lesbian couples and those from single women, are treated as equivalent family structures since both lack a father figure. However, single women shoulder the responsibility for the child's upbringing alone, whereas in lesbian couples partners share this responsibility. Some of the objections raised to alternative types of parenthood therefore refer more to women not having a partner at all. Since 1981, 94 single women have asked the Fertility Centre of the Free University of Brussels to be considered for AID treatment. Data from this group of alternative requests have been collected. The aim is to draw up a profile of the single women who request AID, and to make a distinction between applicants who were accepted and those who were refused.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Effects of etomidate on ovarian steroidogenesis

The present study investigated the influence of two i.v. induction agents, etomidate (0.25 mg/kg) and thiopental (4 mg/kg) followed by isofluorane anaesthesia on the venous blood concentrations of prolactin (PRL), follicle-stimulating hormone (FSH), 17β-oestradiol (E2), progesterone (P), 17OH-progesterone (17OH-P) and testosterone (T) in 18 patients during laparoscopy for oocyte aspiration for in-vitro fertilization. A sharp decrease in the plasma concentration of E2 (P<0.01), P (P<0.01), 17OH-P (P<0.01) and T (P<0.01) was observed within 10 min after induction of anaesthesia with etomidate, followed by a gradual return to the baseline levels thereafter. No such decrease was noted after the administration of thiopental. These findings suggest that etomidate, which has previously been reported to inhibit adrenocortical steroidogenesis, can also interfere with the endocrine function of the ovary. © 1987 IRL Press Limited.SCOPUS: ar.jinfo:eu-repo/semantics/publishe