RESUMO
We studied a new method of treatment of amyotrophic lateral sclerosis with autologous mesenchymal stem cells. Autologous mesenchymal stem cells were injected intravenously (intact cells) or via lumbar puncture (cells committed to neuronal differentiation). Evaluation of the results of cell therapy after 12-month follow-up revealed slowing down of the disease progression in 10 patients in comparison with the control group consisting of 15 patients. The cell therapy was safe for the patients.
Assuntos
Esclerose Lateral Amiotrófica/terapia , Transplante de Células-Tronco Mesenquimais , Adulto , Idoso , Esclerose Lateral Amiotrófica/metabolismo , Esclerose Lateral Amiotrófica/patologia , Diferenciação Celular , Células Cultivadas , Feminino , Humanos , Masculino , Células-Tronco Mesenquimais/enzimologia , Pessoa de Meia-Idade , Nestina/metabolismo , Fosfopiruvato Hidratase/metabolismo , República de Belarus , Transplante Autólogo , Resultado do TratamentoRESUMO
The efficiency of human bone marrow (BM) mesenchymal stem cell (MSC) transduction with a bicistronic lentivirus vector was estimated, and the stability of transgene expression in genetically modified MSCs was determined. First-passage BM MSCs were capable of efficient transduction with the bicistronic lentivirus vector. The transduction efficiency depended on the multiplicity of infection (MOI), being 64 +/- 6.5 and 88.6 +/- 2.9% at MOI 10 and 20, respectively. The lentivirus transduction efficiency proved independent on the number of passages of a BM MSC culture, and expression of the egfp and dsRed1 transgenes in genetically modified MSCs remained stable for one month of culturing. A comparison showed that the level of egfp and dsRed1 transgene expression was preserved upon hepatogenic differentiation in vitro. The results provide a basis for further development of multigenic modification of human BM MSCs for research and/or therapeutic purposes.
Assuntos
Células da Medula Óssea , Vetores Genéticos , Células-Tronco Mesenquimais , Transdução Genética/métodos , Células da Medula Óssea/citologia , Células da Medula Óssea/metabolismo , Diferenciação Celular , Expressão Gênica , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Células HEK293 , Humanos , Lentivirus/genética , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , TransgenesRESUMO
Conditions for culturing and differentiation of human umbilical blood mononuclear cells in vitro were studied. The growth of mesenchymal stem cells was attained in 31 of 54 (57.4%) umbilical blood samples and morphological and immunophenotypical authenticity of these cells was confirmed. Stimulatory effects of 20% AB(IV) human serum and transforming growth factor-beta (TGF-beta) on the growth of mesenchymal stem cells were demonstrated. Osteogenic cells formed in the presence of differentiation factors ascorbic acid, dexamethasone, and beta-glycerophosphate, while chondrogenic cells developed in the presence of dexamethasone, ascorbic acid, and TGF-beta. Differentiation of mesenchymal stem cells was confirmed by histochemical and molecular genetic tests.
