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
Dans Chinois
| WPRIM
| ID: wpr-357269
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>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Plasmides
/
Activation des lymphocytes
/
Lymphocytes T
/
Transfection
/
Lignée cellulaire
/
Mouvement cellulaire
/
Molécule-1 d'adhérence des cellules vasculaires
/
Petit ARN interférent
/
Interférence par ARN
/
Prolifération cellulaire
Limites du sujet:
Animaux
langue:
Chinois
Texte intégral:
Journal of Experimental Hematology
Année:
2015
Type:
Article
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