In Vitro Expansion And Inmmunoregulatory Effect Of Mesenchymal Stem Cells(MSCs) Derived From Human Bone Marrow

Objectives: Bone marrow-derived mesenchymal stem cells(MSCs) were co-cultured with T cells in the presence of PHA or allo-antigens, in order to study the inmmunoregulatory effect and the immunologic tolerance induced by MSCs, as well as the mechanism to reducing graft-versus-host disease rate. Methods: MSCs were isolated and culture-expanded from human bone marrow, and were identified by the spindle fibroblastic morphology characterization by micro-photograph . The phenotypes of MSCs were detected by flow cytometry (FCM). MSCs were added to PHA induced transformation cultures and one-way mixed lymphocyte cultures with various concentrations. Also the supernatant of MSCs was added to one-way mixed lymphocyte cultures with various concentrations. At different time points, the level of cytokines were tested by enzyme linked immunosorbant assay(ELISA), including IL-2 and INF-y(screted by Th1 cells), IL-4 and IL-10(screted by Th2 cells).Results: Mononuclear cells isolated from bone marrow were inoculated to the flasks, and grew slowly as typically fibroblastic shape in 2 to 4 days; Well-adhered cells as fibroblastic shape grew faster in 7 to 11 days, and had uniform morphology...