Cultivation and Neural Differentiation of Embryonic Cerebrospinal Fluid Treated Adipose Stem Cells on the Scaffold of Amniotic Membrane

Introduction: Adipose stem cells (ASCs) are ideal candidates for cell therapy of neurological disorders. In vitro methods require the use of a variety of growth factors and multi-step protocols to induce neuronal differentiation. This study was aimed to assess the neural differentiation of adipose stem cells in a co-culture system. Material and Methods: ASCs were obtained from male Wistar rats and were characterized, using flow cytometry. Harvested ASCs were cultured on a scaffold prepared from amniotic membrane (AM). Cerebrospinal fluid (CSF) was collected from rat embryos and was added to culture medium for 7 days. Structure of scaffold and cell attachment was assessed through scanning electron microscopy (SEM). Neural differentiation of ASCs in the co-culture system was confirmed with immunofluorescence (IF) staining for β-tubulin III and MAP-2 markers. Results: SEM results confirmed the decellularization of AM and attachment of ASCs on the AM derived scaffold. MTT assay revealed that ASCs proliferated on AM significantly during the 7 days of culture. IF data confirmed that the CSF treated cells were expressed by β-tubulin III and MAP-2 but untreated cells were negative for the expression of neural markers. Conclusion: Cultivation of ASCs on the scaffold and their treatment with CSF induced them into the neural lineage fate in the absence of any chemical inducing factor. This method of co-culture may represent a new method to improve in vitro neural differentiation of ASCs

Effect of vitamin E succinate as a differentiation agent on the efficacy of 5-ALA-PDT on prostate cancer cells in culture

 Photodynamic Therapy (PDT) using 5-aminolevulinic acid (5-ALA)-induced protoporphyrin IX (PpIX) has been considered as a new method for treating neoplasms. However, ALA-PDT is suboptimal for thick tumors. Searching for new approaches, we investigated the effect of adding differentiation therapy (DT) with Vitamin E succinate (VES) to PDT in human prostate LN-CaP-FGC10 cancer cells in vitro. The purpose of DT was to reverse the lack of differentiation in cancer cells and to enhance the effectiveness of ALA- dependent PDT.Three groups of cells were grown on RPMI1640 culture medium supplemented with 10% FBS. The cells included: ALA-PDT cells, which received 0.3 mM ALA for 4 hours at dark, and exposed to 532 nm, 50 mW Nd-YAG laser beam for 3 min; DT+ALA-PDT cells, which received 6 µg/ml VES for 24, 48 and 72 hours, followed by the addition of 0.3 mM ALA for 4 h and exposure to Nd-YAG laser beam for 3 min; control cells which were untreated. After 24 h, the percentage of cell viability was determined by MTT assay. Accumulation of PpIX was measured by spectrophotometry and fluorescent microscopy. Mechanism of induced cell death was investigated via Hoechst staining. The combination of both factors (VES and 5-ALA) lead to a significant increase in cell death after 72 h. Induction of differentiation augmented PpIX accumulation in cells treated with ALA. Elevated intracellular PpIX levels resulted in an enhanced lethal photodynamic sensitization of VES plus ALA-treated cells after 72 h. Apoptotic cell death by both ALA-PDT and VES-ALA-PDT was confirmed by Hoechst staining. Our data suggest that VES used in combination with 5-ALA may provide a new combinatorial approach for treating certain cancers.

The effects of Urtica dioica extract on lipid profile, insulin resistance index and liver histology in polycystic ovary syndrome-induced Wistar rats

Background and aims: Urtica dioica as a medicinal herb due to its anti-inflammatory, antioxidant and hypoglycemic effects, improve type 2 diabetes and decrease inflammation, fibrosis and necrosis as the signs of Nonalcoholic steatohepatitis. Polycystic Ovary Syndrome (PCOS) as the most important factor of infertility has an overlap of 30 to 60 with liver disorders. Hence, the Urtica dioica moderator effect on liver function in PCOS rats was examined. Methods: In this experimental study 144 adult Wistar rats were divided into control, PCOS and nettle-treated groups. The group PCOS was injected subcutaneously 2 mg estradiol valerate, after 60 days and confirmed polycystic, the experimental group was injected intraperitoneally the Urtica dioica extract doses (150,250, 450 mg/kg BW). After 21 days, rats were anesthetized by chloroform; blood samples and livers were collected for histological and serological evaluation. Data were analyzed using instat3 via ANOVA one-way and

Development of Mouse Preantral Follicle after In Vitro Culture in A Medium Containing Melatonin

Objective: Improvements in cancer treatment have allowed more young women to survive. However, many cancer patients suffer from ovarian failure. Cryopreservation is one of the solutions for fertility restoration in these patients. The cryopreservation of isolated follicles is a more attractive approach in the long term. Many endocrine and paracrine factors can stimulate the granulosa cells of preantral follicles to proliferate. Melatonin acts as direct free radical scavenger and indirect antioxidant. In this study, we investigated the direct effects of melatonin on follicle development and oocyte maturation by exposing in vitro cultured mouse vitrified-warmed ovarian follicles to melatonin. Materials and Methods: In an experimental study, preantral follicles with diameter of 150-180 μm were isolated from prepubertal mouse ovaries. Follicles were vitrified and thawed using cryolock method. They were then cultured individually for 7 days in droplets supplemented with 0, 10 and 100 pM melatonin, while ovulation was induced using epidermal growth factor (EGF) and human chorionic gonadotropin (hCG). The survival rate of follicles and nuclear maturation of ovulated oocytes were determined. Results: At the end of culture, significant increases in follicle survival (p0.05). Conclusion: Culture of mouse vitrified-warmed preantral follicles in a medium supplemented with 10 pM melatonin increased the number of surviving follicles

Profound changes in cerebrospinal fluid proteome and metabolic profile are associated with congenital hydrocephalus

From SAGE Publishing via Jisc Publications RouterHistory: received 2021-01-16, rev-recd 2021-07-07, accepted 2021-07-12, epub 2021-08-20Publication status: PublishedThe aetiology of congenital hydrocephalus (cHC) has yet to be resolved. cHC manifests late in rodent gestation, and by 18–22 weeks in human fetuses, coinciding with the start of the major phase of cerebral cortex development. Previously we found that cerebrospinal fluid (CSF) accumulation is associated with compositional changes, folate metabolic impairment and consequential arrest in cortical development. Here, we report a proteomics study on hydrocephalic and normal rat CSF using LC-MSMS and a metabolic pathway analysis to determine the major changes in metabolic and signalling pathways. Non-targeted analysis revealed a proteome transformation across embryonic days 17–20, with the largest changes between day 19 and 20. This provides evidence for a physiological shift in CSF composition and identifies some of the molecular mechanisms unleashed during the onset of cHC. Top molecular regulators that may control the shift in the CSF metabolic signature are also predicted, with potential key biomarkers proposed for early detection of these changes that might be used to develop targeted early therapies for this condition. This study confirms previous findings of a folate metabolic imbalance as well as providing more in depth metabolic analysis and understanding of cHC CSF