ABSTRACT
This study was purposed to investigate the effects of mesenchymal stem cells (MSC) on lymphocyte proliferation of sensitized mice in vitro and their action manner so as to more understand the possible mechanisms of bone marrow-derived MSC action on the engraftment of hematopoietic stem/progenitor cells in sensitized mice. Bone marrow-derived MSC were cultured by adherent culture method, the MSC or culture supernatant were used as immune cells or immunologic factor, the spleen lymphocytes of sensitized mice were used as effector cells. The phytohemagglutinin (PHA) was used to stimulate the lymphocyte proliferation, the MTT method was used to detect the mixed lymphocyte reaction. The results showed that bone marrow-derived MSC could inhibit the lymphocyte proliferation of sensitized mice, the MSC cultured supernatant also exhibit this effect. Moreover, the inhibitory effect of MSC supernatent increased along with the increase of cell ratio and concentration, while ratio of these two kind of cells was 1:1, the inhibitory effect was the highest (p < 0.05). It is concluded that the MSC can suppress lymphocyte proliferation of sensitized mice in vitro through cell-cell direct contact or cell-cell indirect interaction.
Subject(s)
Animals , Male , Mice , Bone Marrow Cells , Cell Biology , Allergy and Immunology , Cell Proliferation , Cells, Cultured , Lymphocyte Activation , Allergy and Immunology , Mesenchymal Stem Cells , Cell Biology , Allergy and Immunology , Mice, Inbred BALB C , Mice, Inbred C57BL , Spleen , Cell Biology , Allergy and ImmunologyABSTRACT
AIM:Targeting of focal adhesion kinase (FAK) gene,we aim to construct FAK shRNA lentiviral vector and to identify its function on the growth of leukemic cells.METHODS:FAK shRNA was chemically synthesized,and inserted into a GFP-lentiviral plasmid through molecular biology methods.After packaged and concentrated,the lentiviral-FAK-shRNA-vector was transduced into a human leukemic cell line.FAK gene expression was detected by reverse transcriptional PCR and Western blotting.Cell apoptosis was measured by Annexin V labeling.RESULTS:The results showed that FAK shRNA was successfully inserted into the lentival vector,and the infection efficiency varied from 10% to 25%.Compared to the control vector (lentival vector without FAK shRNA),FAK shRNA inhibited the expression of FAK mRNA and protein by 40% and 70%,respectively.Moreover,the results of apoptosis experiment showed that the percentages of Annexin V+ cells in control vector group and FAK shRNA group were (4.19 ? 0.36) % and (8.48 ? 0.58) % respectively,the difference was statistically significant (P