Ulrika Selleskog

Re-analysis of 166 embryos not transferred after PGS with advanced reproductive maternal age as indication

BACKGROUND In a randomized controlled study aiming to test the effectiveness of preimplantation genetic screening (PGS) in women of advanced maternal age, embryos diagnosed as chromosomally abnormal and those with no diagnosis were fixed for reanalysis. The aim of this study was to determine how well the chromosomal constitution of one biopsied blastomere reflects the status of the entire embryo. METHODS One hundred and seventy-three embryos diagnosed as chromosomally abnormal, 22 with no PGS result and four degenerated embryos originally diagnosed as normal were fixed and reanalysed by fluorescence in situ hybridization. RESULTS In total, 199 embryos were fixed, of which 166 were successfully reanalysed. One hundred and sixty embryos were found to be chromosomally abnormal; 48 of the reanalysed embryos with an initial diagnosis (149) had at least one cell with exactly the same chromosomal constitution shown in the first PGS analysis (34.2%). The reanalysis confirmed the initial overall chromosomally abnormal status of the embryo in 95.9% of the cases. Of all chromosomally abnormal embryos, 4.1% were diagnosed as false positive. The risk for false negative rate was at least 4.1%. CONCLUSIONS PGS seems to be a good method for selecting against chromosomally abnormal embryos but not for determining an embryos exact chromosomal constitution.

Conventional morphology performs better than morphokinetics for prediction of live birth after day 2 transfer

Numerous studies have reported on the potential value of time-lapse variables for prediction of embryo viability. However, these variables have not been evaluated in combination with conventional morphological grading and patient characteristics. The aim of this study was to assess the ability of patient characteristics and embryo morphology together with morphokinetic variables to predict live birth after day 2 transfer. This retrospective analysis included 207 transferred embryos from 199 couples cultured in a time-lapse system up to day 2 of development. Good prediction of live birth or ranking of embryos with respect to live birth potential was achieved with early cleavage combined with fragmentation grade at 43-45 h. These variables were selected as the strongest predictors of live birth, as assessed by stepwise logistic regression, and additional inclusion of morphokinetic variables did not improve the model significantly. Also, neither logistic regression models nor classification tree models with morphokinetic variables were able to achieve equally good prediction of live birth, as measured by AUC on an external data set not used for model development. In conclusion, for fresh day 2 transfers early cleavage in combination with fragmentation grade at 43-45 h should be considered when selecting between good quality embryos.

Regulation of androgen production in cultured human thecal cells by insulin-like growth factor I and insulin**Supported by grant no. 5987 from the Swedish Medical Research Council, Swedish Medical Society, The Medical Society of Göteborg, Kabi-Pharmacia Ltd., Nordisk Insulin Foundation, Hjalmar Svenssons Research Foundation, Swedish Society for Medical Research, and the University of Göteborg, Göteborg, Sweden.

OBJECTIVES To investigate if human thecal cells contain messenger ribonucleic acid (RNA) encoding insulin-like growth factor I (IGF-I) and insulin receptors and if IGF-I and insulin could stimulate androgen production in thecal cells. DESIGN Poly-adenine+ RNA was extracted from fresh thecal tissue, and the expression of the genes encoding insulin and IGF-I receptors were analyzed. Isolated thecal cells were cultured 4 to 6 days with and without hormones. SETTING Procedures were performed in a university laboratory. PATIENTS Eight women in the follicular phase of natural cycles were undergoing gynecological laparotomy for reasons unrelated to ovarian pathology. The leading follicle(s) was excised, and dispersed cells of the theca interna layer were isolated through combined mechanical and enzymatic techniques. INTERVENTIONS Luteinizing hormone (LH), IGF-I, and insulin were added to the cell cultures. MAIN OUTCOME MEASURE The expression of IGF-I receptor and insulin receptor transcripts were analyzed by Northern blot. Medium levels of androstenedione and testosterone were measured by radioimmunoassay. RESULTS In the separated thecal tissue both IGF-I receptor and insulin-receptor transcripts were detected. Insulin-like growth factor I and insulin potentiated LH-induced androgen secretion while having less pronounced effects on basal androgen production. CONCLUSION The present study demonstrates that both insulin and IGF-I receptor genes are expressed and that insulin and IGF-I can stimulate steroid production in human thecal cells. The study provides further support for the hypothesis that IGF-I and insulin may be involved both in physiological regulation of ovarian function as well as in its pathophysiology.