ABSTRACT
Spermatogonial stem cells [SSCs] maintain spermatogenesis throughout life in the male. Maintenance of SSCs and induction of spermiogenesis in vitro may provide a therapeutic strategy to treat male infertility. This study investigated in vitro differentiation of mouse SSCs in presence or absence of Sertoli cells, hormones and vitamins. Spermatogonial populations were enriched from testes of 4-6 week old males by magnetic activated cell sorting and anti-Thy-1 antibody. Sertoli cells isolated from 6-8 week old testes were enriched using lectin-DSA-coated plates. Isolated SSCs were cultured in the presence of Leukemia inhibitory factor [LIF] for 7 days in gelatin-coated dishes, then dissociated and cultured for 7 days in media lacking LIF in the presence or absence of Sertoli cells, with or without FSH, testosterone and vitamins. After one week, the effects of Sertoli cells +/- supplementary media on SSC differentiation was evaluated by microscopy and expression of meiotic and postmeiotic transcripts using RT-PCR. SSC colonies had limited development after LIF removal alone, exhibiting low expression of meiotic [Scp3, Th2b] but not postmeiotic transcript, and loss of Stra8 and Dazl expression. SSCs co-cultured with Sertoli cells, hormones and vitamins developed spermatid-like cells expressing postmeiotic markers [TP1, TP2, Prm1] at levels over 2-fold higher than Sertoli cells or hormone/vitamins alone. Our present SSC-Sertoli co-culture provides conditions that may allow efficient in vitro differentiation of SSCs for the treatment of male infertility
ABSTRACT
Infants are very sensitive and special attention is needed to take care of their physical and mental health. Some studies showed that massage by parents can improve infants' sleep quality. The aim of this study was to compare the effect of massage with sunflower oil or sesame oil on infants' crying and sleep times. This triple-blinded clinical trial was conducted on 120 infants who were 10-15 days old, full-term, single, exclusively breastfed and with no history of hospitalization. The infants were assigned randomly into three groups of sunflower oil massage, sesame oil massage and massage with no oils. Mothers administered 15 minutes of massage to their infants twice per day [morning and afternoon] for 28 days. Times of crying and sleep were measured by a parents' information form at baseline, and at the end of the first, second, third, and fourth weeks of the study. Data were analyzed using the repeated measures ANOVA. Within four weeks of following, decreasing pattern of infants crying time was not significant [P=0.18]; however, the mean of crying time between the three groups was significant [P=0.007]. The interaction effect of crying time and groups was not significant [P=0.18]. Increasing pattern of infant sleep time was statistically significant in the fourth weeks of the following [P<0.001]; however, the mean of sleep time between three groups was not significant [P<0.56]. The interaction effect of sleeping time and groups was not significant [P<0.10]. Since there were not statistical significant differences between the infants' crying and sleep times between the three groups, and despite the fact that increasing pattern of infants sleep time is a normal pattern, it is not possible to have a proper conclusion