[ influence of cerebrospinal fluid accompanied by retinoic acid on differentiation of bone marrow mesenchymal stem cells into neuron-like cells in vitro]

Objective: Cerebrospinal fluid [CSF] has a broad range of molecules and neurotrophic factors essential for neurogenesis. Bone marrow mesenchymal stem cells [BMSCs] are multipotent stem cells that can differentiate into the cells with neural-like phenotype under the induction of appropriate growth factors. According to the significant role of retinoic acid [RA] in neurogenesis, this study aims to differentiate BMSCs into neuronlike cells using CSF, RA, and the combination of CSF and RA

Methods: Rat BMSCs were isolated and characterized. The CSF was prepared from the cisterna magna of 19-day-old Wistar rat embryos. The BMSCs were induced by either 5% CSF [CSF group], 10-6 microM RA [RA group], or CSF plus RA [CSR group] for 12 days. Morphology of differentiated cells was examined by inverted microscope and axonal outgrowth measured using Image J software. In addition, the expression of neural-specific markers [Nestin and MAP-2] was examined by immunocytochemistry

Results: We observed specific-neuronal morphology in the differentiated cells. The maximum axon length was seen in the CSR group on the 12th day of induction. Immunocytochemistry results showed that the neural progenitor marker [Nestin] was expressed in all treated groups. However, MAP-2, as a mature neural marker, was only expressed in the CSR group

Conclusion: The findings suggest that CSF accompanied RA lead to differentiation of cells with neuronal and glial phenotypes from BMSCs in vitro

[ effect of Honey bee venom on induction of differentiation of k562 cell line to erythroid linage]

Complementary medicine uses bee venom [BV] to treat several diseases, including arthritis and skin diseases. BV contains mellitin, phospholipase A2, apamin and several other bioactive substances. According to the venom compounds, the purpose of this study was to examine the effects of BV on differentiation of K562cell line. In this experimental study, K562cells were treated with different doses of BV in different durations. BV toxicity was evaluated by MTT assay. Benzidine staining was used to investigate the effects of BV on K562 cell differentiation toward the erythroid line. In order to determine the type of cell death, annexin-V gene expression was analyzed by flow cytometry. Colony assay was used to measure BV ability in inhibiting colony formation. Morphological changes in the cells undergone treatment with BV were evaluated by wright-giemsa staining. MTT assay showed that bee venom with concentrations of 5.5-6 microg/ml and 3.5-4.5microg/ml result in 505 cell death in 24h and 48h, respectively. Morphological examination and benzidine staining showed that lower doses in longer period induce differentiation in these cells. Flow cytometry data showed significantly increased in annexin-V gene expression in cells which were treated with bee venom for 24 h. Colony assay demonstrated that the concentration of 1 microg/ml of BV results in 50% reduction in colony formation

[Induction of differentiation of stem cells derived from adipose tissue to ocular lens fiber cells]

Presumably, stem cells derived from mouse adipose tissue differentiate into lens fiber cells under induction by vitreous humor factors. Cataract is one of the most common ocular diseases that is highly treatable by replacing the ocular lens with artificial ones. The objective of this study was to find the natural lens replacements instead of the artificial ones. In this experimental study, stem cells from adipose tissue were obtained from inguinal fat pads from NMRI mouse. To demonstrate stemness potential of these cells, we used anti-OCT4, as stromal stemness markers with immunocytochemistry methods. This experiment took place for 14 days with different dosages of bovine vitreous humor. Express of ocular lens fiber cells markers [alpha crystalline] were detected by immunocytochemistry in experimental and control groups. All murine stem cells were positive to OCT4 marker which is used for stromal cells. Morphological studies showed that cells induced with 40% vitreous humor in culture media were locally longer and more aligned in parallel compared to control group cells. Also, the fiber like cells had large nuclei with multiple nucleoli that showed increase in production of proteins in those cells. The proof of the induction of these stem cells to lens fiber like-cells is the positive response of induced cells for crystalline markers in media with 40% vitreous humor by ANOVA test. Based on morphological appearance and positive response of experimental group to specific markers of lens fiber cells, it can be concluded that stem cells derived from mouse adipose tissue differentiate into lens fiber cells by treating them with vitreous humor

effect of cholestasis on rewarding and exploratory behaviors induced by opioidergic and dopaminergic agents in mice

Several investigations have indicated that cholestasis decreases opioid receptor expression in the brain following increased opioidergic neurotransmission. The opioidergic system plays an important role in regulation of reward circuits that may be produced via dopamine-dependent mechanisms. It has been suggested that the dopaminergic system of the nucleus accumbens is necessary in conditioned place preference [CPP]. The aim of this study is, therefore, to test if cholestasis can alter the reward system and the involvement of opioidergic and dopaminergic systems in this phenomenon. We used CPP and hole-board paradigms to measure the reward effect and exploratory behaviors, respectively, in mice. Cholestasis was induced by ligation of the main bile duct, using two ligatures and transecting the duct between them [BDL mice]. The data showed that morphine [1 and 2 mg/kg], sulpiride [80 mg/kg] and SKF38393 [20 mg/kg] produced CPP, while naloxone [1 mg/kg] and SCH23390 [1 mg/kg] produced conditioned place aversion [CPA], whereas quinpirole had no effect in sham-operated mice. However, morphine [2 mg/kg, i.p.], sulpiride [40 mg/kg] and SKF38393 [10 mg/kg] induced CPP in BDL mice compared to sham-operated mice. Naloxone- or SCH23390-induced CPA was reduced in BDL mice compared with the respective sham-operated mice. Quinpirole tended to induce aversion in BDL mice which was, however, not significant. In addition, quinpirole 1 mg/kg] and SCH23390 [1 mg/kg] increased head-dip exploratory behavior, whereas naloxone [2 mg/kg] caused a decrease in head-dip exploratory behavior in sham-operated mice. Morphine [2 mg/kg], SCH23390 [1 mg/kg] and quinpirole [0.25 and 0.5 mg/kg] induced anxiogenic-like behavior in BDL mice. It can be concluded that cholestasis differentially alters the reward effects of opioidergic and dopaminergic agents

Derivation of adipocytes from human endometrial stem cells [EnSCs]

Due to increasing clinical demand for adipose tissue, a suitable cell for reconstructive adipose tissue constructs is needed. In this study, we investigated the ability of Human Endometrial-derived stem cells [EnSCs] as a new source of mesenchymal stem cells to differentiate into adipocytes. EnSCs are the abundant and easy available source with no immunological response, for cell replacement therapy. Single-cell suspensions of EnSCs were obtained from endometrial tissues from 10 women experiencing normal menstrual cycles, and were cultured at clonal density [10 cells/cm[2]] or limiting dilution. Endometrial mesenchymal stem cell markers were examined flow cytometry. These cells were treated with adipogenicinducing medium for 28 days. The adipogenic differentiation of the EnSC was assessed by cellular morphology and further confirmed by Oil Red O staining and RTPCR. The BM-MSC differentiated into adipocytes in the presence of adipogenic stimuli for 3 weeks. The flow cytometric analysis showed that the cells were positive for CD90, CD105, CD146 and were negative for CD31, CD34.We showed that the key adipocytes marker PPARa was expressed in mRNA level after 28 days post treatment [PT]. According to our finding, it can be concluded that EnSCs represent a useful in vitro model for human adipogenesis, and provide opportunities to study the stages prior to commitment to the adipocyte lineage