ABSTRACT
Background: Mesenchymal stem cells (MSCs) are deemed as potential new therapeutic agents for infertility treatment and adipose tissue (AT) becomes a potential MSCs source. To direct MSCs through the differentiation process properly, an environment comparable to the in vivo niche might be indispensable. Objective: This study aims to differentiate human AT-derived MScs (hAD-MScs) into male germ-like cells in vitro using a combination of rabbit Sertoli cells conditioned medium (SCCM), bone morphogenetic protein 4, and retinoic acid. Materials and Methods: MScs were isolated from human ATs of fertile and infertile donors. The verified MScs were differentiated using a 2-step protocol; the first step included 20 ng/ml bone morphogenetic protein 4 treatment. The second step was performed utilizing 1 µM retinoic acid and/or SCCM. The morphological changes and the expression of germ cell (GC)-specific markers: octamer-binding transcription factor-4; stimulated by retinoic-acid-8, synaptonemal complex protein-3, andprotamine-1 were assessed in the treated cells using quantitative polymerase chain reaction. Results: Induction of hAD-MScs resulted in the upregulation of GC-specific genes where SCCM treatment showed the highest expression. The synaptonemal complex protein-3andprotamine-1 gene expression was detected after 19 and 26 days of induction, respectively. PRM1 was detected in hAD-MScs cultured in SCCM earlier than in other treated groups. The treated cells became more elongated-like spindles and formed aggregates. Conclusion: hAD-MScs differentiated to GC lineage exhibited the ability to express GC-specific markers under in vitro conditions, and rabbit's Sertoli cells can be used for inducing transdifferentiation of hAD-MScs into germ-like cells.
ABSTRACT
UNLABELLED: Back ground: Careful evaluation of patients and proper treatment with right techniques are essential for successful outcome of assisted reproduction. To obtain satisfactory results, it is necessary to assess ovarian reserve before planning treatment. OBJECTIVE: To evaluate anti-mullerian hormone as a predictor of fertility potential in terms of ovarian reserve and ovarian response reflected by antral follicles and mature oocyte counts in response to menotrophin stimulation in in vitro fertilization (IVF) women from Gaza Strip. MATERIALS AND METHODS: This prospective cohort study consisted of 81 women (mean age 28.7 years) attending IVF at Al-Basma Fertility Center in Gaza City. Blood withdrawal for antimullerian hormone measurement was performed in all the patients and the number of oocytes and embryos were recorded. RESULTS: The total number of retrieved oocytes was inversely associated with age (12.5±4.5, 11.0±5.4 and 6.9±4.7 at age ≤25, 26-35 and >35 years, respectively (F=4.793 and p=0.011). The ovarian response to Menotrophin (FSH 75IU, LH 75 IU) stimulation was better for younger age. There was a significant positive association between ovarian response in terms of total number of oocytes and antimullerian hormone levels. The maximum level of antimullerian hormone was observed in females who achieved positive pregnancies (4.5±2.5 ng/mL) followed by negative pregnancies (2.9±1.8 ng/mL) with significant differences (F=6.862 and p=0.002). Correlation coefficient revealed that the number of mature oocytes showed strong positive correlation with the antimullerian hormone levels (r=0.469, p=0.001). CONCLUSION: Anti-mullerian hormone can be used in IVF programs as a good predictor of ovarian reserve and ovarian response.
