The effect of maternal sleep deprivation on differentiation of mesenchymal stem cells in the presence of neonates brain cerebrospinal fluid of Wistar rats.

OBJECTIVE: Cerebrospinal fluid (CSF) contains proliferation, differentiation and maturation signals that are essential factors for brain development. Due to presence of such factors this fluid has proliferative and differentiation potential. A previous study showed that maternal sleep deprivation (MSD) decrease the number of newborn neurons in development of hippocampus. Also, it impairs hippocampus-dependent spatial learning and memory in the young offspring rat. MSD can change CSF factors. In the present study, the effect of MSD-CSF on differentiation of mesenchymal stem cells to neural cells was examined, and expression of Nestin, Neun, and NeurD1 that are neurogenic markers was investigated. MATERIAL AND METHODS: In this study, bone marrow mesenchymal stem cells were aspirated from the femur and tibia of young male Wistar rats. Then cell suspension was cultured in DMEM medium supplemented with 10 % FBS and 1 % antibiotics. Pregnant rats were subjected to sleep deprivation for 6 h by gentle handling during 4th, 7th, and 18th day of pregnancy. CSF was collected from cisterna magna of neonate rats. CSF was added to culture media with a 5 % ratio (v/v). Then cell viability was determined with MTT assay. Total cellular RNA was extracted, cDNA was synthesized and NeuN, Nestin, NeuroD1 and IL-6 genes were analyzed by Real-time PCR. RESULTS: Real time-PCR analysis showed that expression of Neun and NeurD1gene decreased compared with culture in normal CSF (N-CSF), and also showed that expression of Nestin did not decrease, inflammatory gene (IL-6) showed high expression compared to culture with N-CSF. CONCLUSION: Based on our results, MSD-CSF could inhibit neurogenesis process in mesenchymal stem cells and also, this result suggests that MSD can suppress neural differentiation and decrease of neurogenesis gene expression. Overall these findings suggest that insomnia and sleep loss may active inflammatory responses in the brain and change CSF-markers (Fig. 3, Ref. 34).

Evaluation of Epidermal Neural Crest Stem Cells in Organotypic Spinal Cord Slice Culture Platform.

Among various strategies employed for spinal cord injury, stem cell therapy is a potential treatment. So far, a variety of stem cells have been evaluated in animal models and humans with spinal cord injury, and epidermal neural crest stem cells represent one of the attractive types in this area. Although these multipotent stem cells have been assessed in several spinal cord injury models by independent laboratories, extensive work remains to be done to ascertain whether these cells can safely improve the outcome following human spinal cord injury. Among the models that closely mimic human spinal cord injury, the in vitro model of injury in organotypic spinal cord slice culture has been identified as one of the faithful platforms for injury-related investigations. In this study, green fluorescent protein-expressing stem cells were grafted into injured organotypic spinal cord slice culture and their survival was examined by confocal microscope seven days after transplantation. Data obtained from this preliminary study showed that these stem cells can survive on top of the surface of injured slices, as observed on day seven following their transplantation. This result revealed that this in vitro model of injury can be considered as a suitable context for further evaluation of epidermal neural crest stem cells before their application in large animals.

Bee venom treatment reduced C-reactive protein and improved follicle quality in a rat model of estradiol valerate-induced polycystic ovarian syndrome

Polycystic ovarian syndrome (PCOS) is a low grade inflammatory disease characterized by hyperandrogenemia and chronic anovulation. C-reactive protein (CRP), released by adipocytes, plays a key role in PCOS. Apis mellifera honeybee venom (HBV) contains a variety of biologically active components with various pharmaceutical properties. This study was designed to assess the possibility of HBV application as an anti-inflammatory therapeutic agent. To induce PCOS, 1 mg/100 g body weight estradiol valerate (EV) was subcutaneously (SC) injected into eight-week-old rats. After 60 days, 0.5 mg/kg HBV was administered SC for 14 consecutive days, and the results of PCOS treatment were investigated. Rats were then anesthetized with chloroform, and their ovaries and livers were surgically removed to determine histomorphometrical changes. Testosterone and 17-β-estradiol were detected by chemiluminescence immunoassay. In order to detect serum CRP, ELISA kit was used in three groups of EV-induced PCOS, HBV-treated PCOS and control animals. Thickness of the theca layer, number of cysts and the level of serum CRP significantly decreased in HBV group in comparison with PCOS group. Moreover, corpus luteum, as a sign of ovulation, was observed in HBV-treated ovaries which were absent in PCOS group. Our results suggest that the beneficial effect of HBV may be mediated through its inhibitory effect on serum CRP levels.(AU)