RESUMO
Expression of transgenes in neurons and stromal/mesenchymal stem cells (MSC) can greatly enhance their therapeutic potential. In transfection experiments, we studied properties of linear and branched (dendrimers) polycations as transgene delivery vehicles. Linear polyethyleneimine transfected neurons, but was ineffective in MSC. Polyamidoamine dendrimers showed greater transfection efficiency and mean GFP fluorescence intensity compared to phosphorus dendrimers of the same (4th) generation. Expression of neurotrophic factor BDNF in MSC transfected with polyamidoamine dendrimers was also by more than 10 times higher.
Assuntos
Dendrímeros/química , Células-Tronco Mesenquimais/metabolismo , Neurônios/metabolismo , Poliaminas/química , Transfecção/métodos , Fator Neurotrófico Derivado do Encéfalo/genética , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Expressão Gênica , Genes Reporter , Proteínas de Fluorescência Verde , Humanos , Células-Tronco Mesenquimais/citologia , Neurônios/citologia , Plasmídeos/genética , Polieletrólitos , Polietilenoimina/química , TransgenesRESUMO
We studied the potential of neurogenic induction of human bone marrow mesenchymal stem cells in fibrin-based 3D matrix. The best results were obtained after incubation of mesenchymal stem cells in fibrin 3D matrix in the presence of neurogenic medium containing EGF and bFGF growth factors. Under these conditions, most cells formed numerous branching processes and expressed neural cell marker ßIII-tubulin. Optimal combination of 3D matrix and neurogenic factors of the medium can provide new insight into neurogenic potential of mesenchymal stem cells, which can be used for the therapy of neural traumas and neurodegenerative diseases.
Assuntos
Técnicas de Cultura de Células/métodos , Diferenciação Celular , Células-Tronco Mesenquimais/metabolismo , Neurônios/citologia , Células da Medula Óssea/metabolismo , Meios de Cultura , Fator de Crescimento Epidérmico/farmacologia , Fibrina , Fatores de Crescimento de Fibroblastos/farmacologia , Humanos , Alicerces Teciduais , Tubulina (Proteína)/metabolismoRESUMO
Human bone marrow MSC cultured in neurogenic medium containing EGF and FGFbeta demonstrated alteration of the phenotype and expression of neuronal precursor/early neuron markers nestin and NSE. Signals of expression of neuronal and oligodendroglial markers MAP-2, dm-20, and MBP were detected after prolongation of incubation in neurogenic medium to 2 weeks. Cells with neuronal morphology were immunopositive to early neuronal marker beta-III-tubulin. Replacement of neurogenic medium for alpha-MEM with 10% fetal calf serum induced reversion of the phenotype to that typical for human MSC. This indicates high plasticity of the phenotype and expression profile of neuronal markers in MSC cultured under neurogenic conditions or possibility of dedifferentiation of MSC reaching the stage of neuronal precursors/early neurons.
Assuntos
Células da Medula Óssea/fisiologia , Células-Tronco Mesenquimais/fisiologia , Neurônios/fisiologia , Diferenciação Celular/fisiologia , Meios de Cultura , Citometria de Fluxo , Fluorescência , Humanos , Imuno-Histoquímica , Neurogênese/fisiologia , Fenótipo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
We compared the efficiency of transduction with lentivectors based on HIV-1 and adeno-associated viruses serotype 2 and stability of transgene expression in human mesenchymal bone marrow stem cells.
