ABSTRACT
Objective: cerebrospinal fluid [CSF] plays an important role in cortical development during the fetal stages. Embryonic CSF [E-CSF] consists of numerous neurotrophic and growth factors that regulate neurogenesis, differentiation, and proliferation. Mesenchymal stem cells [MSCs] are multi-potential stem cells that can differentiate into mesenchymal and non-mesenchymal cells, including neural cells. This study evaluates the prenatal and postnatal effects of CSF on proliferation and neural differentiation of bone marrow MSCs [BM-MSCs] at gestational ages E19, E20, and the first day after birth [P1]
Materials and Methods: in this experimental study, we confirmed the mesenchymal nature of BM-MSCs according to their adherence properties and surface markers [CD44, CD73 and CD45]. The multi-potential characteristics of BMMSCs were verified by assessments of the osteogenic and adipogenic potentials of these cells. Under appropriate in vitro conditions, the BM-MSCs cultures were incubated with and without additional pre- and postnatal CSF. The MTT assay was used to quantify cellular proliferation and viability. Immunocytochemistry was used to study the expression of MAP-2 and beta-III tubulin in the BM-MSCs. We used ImageJ software to measure the length of the neurites in the cultured cells
Results: BM-MSCs differentiated into neuronal cell types when exposed to basic fibroblast growth factor [b-FGF]. Viability and proliferation of the BM-MSCs conditioned with E19, E20, and P1 CSF increased compared to the control group. We observed significantly elevated neural differentiation of the BM-MSCS cultured in the CSF-supplemented medium from E19 compared to cultures conditioned with E20 and P1 CSF group
Conclusion: the results have confirmed that E19, E20, and P1 CSF could induce proliferation and differentiation of BM-MSCs though they are age dependent factors. The presented data support a significant, conductive role of CSF components in neuronal survival, proliferation, and differentiation
ABSTRACT
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 micro g/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