Preliminary study of chitosan/pcDNA-EGFP-TGFbeta1 nanoparticles used in the transfection of synovial-derived mesenchymal stem cells.
- Author:
Bin LING
;
Hui LIU
;
Yishan LIU
;
Abass KEREMU
;
Zhongcheng GONG
;
Mei HU
;
Xiaopeng YIN
;
Bo SHAO
;
Zhaoquan LIN
- Publication Type:Journal Article
- MeSH:
Animals;
Chitosan;
chemistry;
Genetic Vectors;
Green Fluorescent Proteins;
genetics;
Mesenchymal Stromal Cells;
cytology;
Microscopy, Electron, Transmission;
Microscopy, Fluorescence;
Nanoparticles;
chemistry;
Rabbits;
Transfection;
Transforming Growth Factor beta1;
genetics
- From:
Journal of Biomedical Engineering
2013;30(6):1260-1264
- CountryChina
- Language:Chinese
-
Abstract:
The objective of this study is to explore the application possibility of chitosan/pcDNA-EGFP-TGFPbeta1 nanoparticles in the transfection of synovial-derived mesenchymal stem cells (SDMSCs). Chitosan/pcDNA-EGFP-TGFbeta1 nanoparticles were fabricated through method of ionic crosslinking. The SDMSCs were harvested from rabbit joints and cultured to passage 3. The SDMSCs were then transfected with chitosan/pcDNA-EGFP-TGFbeta1 nanoparticles. Scanning electronic microscope (SEM) was employed to detect the shape and diameter of the nanoparticles. The transfected SDMSCs were examined under the fluorescence microscope and detected through the flow cytometry (FCM). The SEM examination showed that the contour of the fabricated chitosan/pcDNA-EGFP-TGFbeta1 nanoparticles was round and its average diameter was 50 nm. After being cultured for 48 h, the SDMSCs transfected by chitosan/pcDNA-EGFP-TGFbeta1 nanoparticles could be detected under the fluorescence microscope, and the live SDMSCs could also be examined through FCM. The transfection rate was 8% - 10%. Therefore, it suggested that the chitosan/pcDNA-EGFP-TGFbeta1 nanoparticles fabricated through the method of ionic crosslinking could transfect the SDMSCs, but the transfection rate should be improved.
