The main problem with using MSC (mesenchymal stem cells) to treat the deficient diseases of the central nervous system is the low cell survival rate after the transplant procedure and their low ability to spontaneously differentiate into functional neurons. The aim of this study was to investigate the effects of genetically modifying MSC. A co-overexpression of two genes was performed: BCLXL was supposed to increase the resistance of the cells to the toxic agents and BDNF was supposed to direct cells into the neuronal differentiation pathway. As a result, it was possible to obtain the functional overexpression of the BCLXL and BDNF genes. These cells had an increased resistance to apoptosis-inducing toxicants (staurosporine, doxorubicin and H2O2). At the same time, the genes of the neuronal pathway (CHAT, TPH1) were overexpressed. The genetically modified MSC increased the survival rate under toxic conditions, which increased the chance of surviving a transplant procedure. The obtained cells can be treated as neural cell progenitors, which makes them a universal material that can be used in various disease models. The production of neurotransmitters suggests that cells transplanted into the brain and subjected to the additional influence of the brain’s microenvironment, will be able to form synapses and become functional neurons.
One of the problems with using MSCs (mesenchymal stem cells) to treat different neurodegenerative diseases of the central nervous system is their low ability to spontaneously differentiate into functional neurons. The aim of this study was to investigate how the co-overexpression of the BCL and BDNF genes affects the ability of genetically modified MSCs to differentiate into GABA-ergic neurons. A co-overexpression of two genes was performed, one of which, BCL, was supposed to increase the resistance of the cells to the toxic agents in the brain environment. The second one, BDNF, was supposed to direct the cells onto the neuronal differentiation pathway. As a result, the co-overexpression of both BCL2 + BDNF and BCLXL + BDNF caused an increase in the MAP2 gene expression level (a marker of the neuronal pathway) and the SYP gene that is associated with synaptogenesis. In both cases, approximately 18% of the genetically modified and then differentiated cells exhibited the presence of the GAD protein, which is characteristic of GABA-ergic neurons. Despite the presence of GAD, after both modifications, only the BCL2 and BDNF co-overexpression correlated with the ability of the modified cells to release gamma-aminobutyric acid (GABA) after depolarization. Our study identified a novel model of genetically engineered MSCs that can be used as a tool to deliver the antiapoptotic proteins (BCL) and neurotrophic factor (BDNF) directly into the brain microenvironment. Additionally, in the investigated model, the genetically modified MSCs could easily differentiate into functional GABA-ergic neurons and, moreover, due to the secreted BCL and BDNF, promote endogenous neuronal growth and encourage synaptic connections between neurons.
Summary Introduction Flavonoids are widely distributed in the wild. They constitute a large group of compounds that have a beneficial effect on the human body. Apigenin and kaempferol, which belong to the flavone subgroup, have, inter alia, an antitumor effect. The influence of these compounds on the survival of stem cells in a toxic environment has not yet been studied. Objective The aim of the study was to evaluate the effect of selected concentrations of apigenin and kaempferol on the survival of mesenchymal stem cells (MSC) in the presence of a cell-death inducer – staurosporine. Methods Mesenchymal stem cells that were obtained from the Wharton’s jelly of umbilical cords were used for the research. In the first stage, the MSC were treated with apigenin at concentrations of 1.2, 12.5, 25, 50 and 100 µM/ml and kaempferol at concentrations of 1.2, 12.5, 25, 50 and 100 µM/ml. In the next stage, the effect of increased concentrations of 0.1, 0.5 and 1 µM/ml of staurosporine on MSC was examined. The key stage of the experiment was investigating the interaction between the selected concentrations of apigenin (12.5 and 50 µM/ml) and kaempferol (12.5 and µM/ml) on MSC in the presence of staurosporine at a concentration of 1 µM/ml, which had the highest toxicity. Results Both apigenin and kaempferol significantly increased the cytotoxic features of staurosporine on the MSC culture.
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