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1.
Journal of Experimental Hematology ; (6): 858-864, 2016.
Article in Chinese | WPRIM | ID: wpr-246854

ABSTRACT

<p><b>OBJECTIVE</b>To compare the biological characteristics and immunosuppressive activity between human amniotic mesenchymal stem cells (hAMSC) and human bone marrow mesenchymal stem cells (hBMMSC).</p><p><b>METHODS</b>MSC from human amnion and bone marrow were isolated using enzymatic digestion and Ficoll-Hypaque density gradients, respectively. Their biological characteristics were compared by morphology, cell growth curves, cell cycle profile analysis, immunophenotype and immunofluorescence assay. Their immunosuppressive activities were studied on total activated T-cells with phytohemagglutinin (PHA-PBMSC). An in vitro co-culture was performed to compared the lymphocyte proliferation and the supernatant level of IFN-γ were measured by CCK-8 method and ELISA, respectively.</p><p><b>RESULTS</b>Both hAMSC and hBMMSC demonstrated fibroblast-like morphology. The hAMSC were able to be amplified for at least 15 passages, while the hBMMSC only for 6-7 passages. There was no significant difference in the proportion of G2/M phase cells of the 2 cells types (P>0.05). By FACS analysis for immunophenotype, both MSC were shown to be positive for CD105, CD90, CD73 and negative for CD34, CD45, CD11b, CD19, HLA-DR, but hAMSC were positive for Oct-3/4, which was in contrast to hBMMSC. Both of them expressed vimentin. Both the cells exhibited a inhibitory role on the lymphocyte proliferation induced by PHA in co-culture conditions, that was increased with the increase MSC proportion and both the suppressing effecs were enhanced. The supernatant IFN-γ levels of hAMSC co-cultured with lymphocyte at a ratio of 1:1 after 72 hours were measured by ELISA, and the level of IFN-γ was significantly lower than that in the same co-culture system of hBMMSC. In contrast to the IFN-γ in the PHA-stimulated group, the IFN-γ level in both co-culture groups was significantly lower.</p><p><b>CONCLUSION</b>MSC from amnion displayed a higher proliferative capacity and stem cell properties, compared with hBMMSC. Both MSC can inhibit lymphocyte proliferation and suppress IFN-γ secretion induced by PHA in vitro.</p>


Subject(s)
Humans , Amnion , Cell Biology , Bone Marrow Cells , Cell Biology , Cell Proliferation , Cells, Cultured , Coculture Techniques , Hematopoietic Stem Cells , Cell Biology , Immunophenotyping , Immunosuppression Therapy , Lymphocyte Activation , Mesenchymal Stem Cells , Cell Biology , T-Lymphocytes , Cell Biology
2.
Journal of Southern Medical University ; (12): 1422-1427, 2015.
Article in Chinese | WPRIM | ID: wpr-333611

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of simulated microgravity on erythroid differentiation of K562 cells and explore the possible mechanism.</p><p><b>METHODS</b>The fourth generation rotating cell culture system was used to generate the simulated microgravity environment. Benzidine staining was used to evaluate the cell inhibition rate, and real-time quantitative PCR (qRT-PCR) was used to detect GATA-1, GATA-2, Ets-1, F-actin, β-Tubulin and vimentin mRNA expressions. The changes of cytoskeleton were observed by fluorescence microscopy, and Western blotting was employed to assay F-actin, β-tubulin and vimentin protein expression levels.</p><p><b>RESULTS</b>Benzidine staining showed that simulated microgravity inhibited erythroid differentiation of K562 cells. K562 cells treated with Hemin presented with increased mRNA expression of GATA-1 and reduced GATA-2 and Ets-1 mRNA expressions. Simulated microgravity treatment of the cells resulted in down-regulated GATA-1, F-actin, β-tubulin and vimentin mRNA expressions and up-regulated mRNA expressions of GATA-2 and Ets-1, and reduced F-actin, β-tubulin and vimentin protein expressions. Exposure to simulated microgravity caused decreased fluorescence intensities of cytoskeletal filament F-actin, β-tubulin and vimentin in the cells.</p><p><b>CONCLUSION</b>Simulated microgravity inhibits erythroid differentiation of K562 cells possibly by causing cytoskeleton damages to result in down-regulation of GATA-1 and up-regulation of GATA-2 and Ets-1 expressions.</p>


Subject(s)
Humans , Actins , Metabolism , Cell Differentiation , Down-Regulation , GATA1 Transcription Factor , Metabolism , GATA2 Transcription Factor , Metabolism , Hemin , Pharmacology , K562 Cells , Proto-Oncogene Protein c-ets-1 , Metabolism , Tubulin , Metabolism , Up-Regulation , Vimentin , Metabolism , Weightlessness Simulation
3.
Journal of Southern Medical University ; (12): 129-132, 2015.
Article in Chinese | WPRIM | ID: wpr-239221

ABSTRACT

We report two cases of rituximab (RTX)-induced interstitial pneumonia in two lymphoma patients receiving RTX treatment. Interstitial pneumonia was successfully managed in these two cases after a one-week-long intervention with corresponding treatments without affecting further treatment of the primary disease. RTX-induced interstitial pneumonia is characterized by a latent onset with an unclear pathological mechanism and absence of typical symptoms. High-resolution CT scan can provide valuable evidence for early diagnosis of RTX-induced interstitial pneumonia, which might be attributed partially to an increased susceptibility to P. jirovecii and fungal infection due to prolonged RTX treatment.


Subject(s)
Humans , Antibodies, Monoclonal, Murine-Derived , Disease Susceptibility , Lung Diseases, Interstitial , Rituximab , Tomography, X-Ray Computed
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