Neurogenic and mitotic effects of dehydroepiandrosterone on neuronal-competent marrow mesenchymal stem cells.

To establish whether dehydroepiandrosterone (DHEA) as a neurosteroid could enhance the rate of neuronal differentiation in neuronal-competent bone marrow mesenchymal stem cells (BM-MSCs), we added DHEA before and after plating the neurosphere-like aggregates. Flow cytometric analysis of Tubulin-III and Tau positive cells revealed that the percentages of these cells were increased significantly for the two markers following DHEA treatment at both stages. Moreover, Western blot analysis revealed that Tubulin-III protein was strongly induced by DHEA. The expression of neuronal specific genes such as Isl-1, Tubulin III, Pax6 and Nestin was also detected by RT-PCR analysis as well as BrdU incorporation and found to have increased significantly after DHEA induction. In conclusion, these results provide evidence that DHEA can affect neuronal-competent MSCs in inducing the expression of a comprehensive set of genes and proteins that define neuronal cells. DHEA was also able to induce the division of neuronal-competent MSCs, thereby increasing the number of cells with major neuronal characteristics. To our knowledge, this is the first report which shows that DHEA can induce the division and differentiation of MSCs into neurons in vitro and should provide an improved basis for new treatments using MSCs of a wide variety of neurological diseases.

Evaluation of ovulation induction protocols for poor responders undergoing assisted reproduction techniques.

OBJECTIVE: To compare 3 stimulation protocols in poor ovulation responders undergoing in-vitro fertilization (IVF). METHODS: The study was a randomized, prospective clinical trial from June 2003 to July 2004, in Royan Institute, Tehran, Iran. One hundred and fifty-four patients, who had poor responses to ovulation induction in at least one previous IVF attempt, were randomly divided into 3 groups. In the first group, human menopausal gonadotropin (HMG) was administered from day 3 of the cycle at a dose rate of 150 IU/day. In the second group, gonadotropin-releasing hormone (GnRH) agonist was started at a dose rate of 800 microg/day by nasal spray or 500 microg/day subcutaneously in the mid-luteal phase, followed by a standard HMG dose after pituitary down regulation was confirmed. In the third group, clomiphene at a dose rate of 100 mg/day was given from day 3 and HMG from day 6. Our main outcomes were number of mature oocytes, cancellation rate, number of HMG ampoules used and incidence premature luteinizing hormone (LH) surge. RESULTS: There was a high incidence of premature LH surge in all groups except in the GnRH group (p=0.0001) and there were significant differences between groups in HMG requirements (p=0.004). There were no significant differences between groups in number of mature oocytes recovered and cancellation rate. CONCLUSION: Results showed no advantage in the use of GnRH agonist compared to the older regimens of clomiphene plus HMG and HMG alone. The cancellation rate was similar for 3 protocols and HMG requirement was higher with the use of GnRH agonist. The treatment of poor responders in assisted reproductive technologies remains a challenge.