Effect of Knockdown of Vascular Cell Adhesion Molecule-1 on the Immunologic Regulation Capacity of Murine Mesenchymal Stem Cells / 中国实验血液学杂志
Journal of Experimental Hematology
; (6): 796-801, 2015.
Article
en Zh
| WPRIM
| ID: wpr-357269
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To establish the stably lower expression of vascular cell adhesion molecule-1 (VCAM-1) in MSC cell line (C3H10T1/2) by siRNA technology, and explore the effect of knockdown of VCAM-1 on the immunologic regulation capacity of murine MSC.</p><p><b>METHODS</b>The mouse GV118-VCAM-1-RNAi retrovirus vector was constructed by gene recombination technology. The recombinant plasmid was identified by restriction analysis and sequencing, and then the recombinant plasmid GV118-VCAM-1-RNAi was transfected into 293 cells by Lipofectamine, and the supernatant was collected to transfect C3H10T1/2. Moreover, the VCAM-1 lower expression on MSC was evaluated by flow cytometry and fluorescent microscopy. The knockdown VCAM-1 MSC was sorted by flow cytometry. Furthermore, the inhibitory effect of the knockdown VCAM-1 MSC on lymphocyte proliferation was tested by lymphoblast transformation assay (LTT) and mixed lymphocyte reaction assay(MLR).</p><p><b>RESULTS</b>The recombinant retroviral vector of knockdown VCAM-1 (GV118-VCAM-1-RNAi) was successfully constructed and transfected into mouse MSC cell line C3H10T1/2. The knockdown VCAM-1/MSC was obtained by flow cytometric sorting. The LTT and MLR assay showed that the immunosuppressive effect of MSC lower-expressing VCAM-1 dramatically decreased (P<0.05).</p><p><b>CONCLUSION</b>Knockdown VCAM-1 in MSC can significantly down-regulate the inhibitory capability of MSC on the proliferation of T-cells. The data of this study laid an experimental foundation for studying effect of VCAM-1 transfecting into MSC on immune function.</p>
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WPRIM
Asunto principal:
Plásmidos
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Activación de Linfocitos
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Linfocitos T
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Transfección
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Línea Celular
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Movimiento Celular
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Molécula 1 de Adhesión Celular Vascular
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ARN Interferente Pequeño
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Interferencia de ARN
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Proliferación Celular
Límite:
Animals
Idioma:
Zh
Revista:
Journal of Experimental Hematology
Año:
2015
Tipo del documento:
Article