Amaxa Nucleofector(TM) nuclear transfection apparatus transfers L1210 cell line.
- Author:
A-Peng YANG
1
;
Jie-Xian MA
;
Wen-Ming QIN
;
Yan-Hui XIE
;
You-Xin JIN
Author Information
1. Department of Hematology, Huashan Hospital, Fudan University, Shanghai 200040, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Cell Line, Tumor;
Cell Nucleus;
genetics;
Cell Survival;
genetics;
Genes, Reporter;
Green Fluorescent Proteins;
genetics;
Liposomes;
Mice;
Transfection;
methods
- From:
Journal of Experimental Hematology
2010;18(6):1604-1608
- CountryChina
- Language:Chinese
-
Abstract:
Mouse L1210 leukemia cell line is widely used as a model in the study of tumorigenesis, as well as the efficacy of chemotherapeutic drugs; however, like other suspension cell lines, the mouse L1210 cell line has lowest transfection efficiency, that many barriers exist to study about the structure, function, as well as metabolism in leukemia cells. This study was aimed to obtain higher transfection efficiency of L1210 cell line to facilitate scientific research. The transfection efficiencies of nucleofector and liposome in L1210 leukemia cells were detected by converted fluorescence microscopy and flow cytometry using EGFP (enhance green fluorescent protein); cell viability was observed by trypan blue exclusion test. The results showed that the transfection efficiency of nucleofector primarily through reporter gene pEGFP by Amaxa Nucleofector(TM) nuclear transfer apparatus was significantly higher than lipofectamine 2000 transfection, furthermore, in the same cell density (2 × 10(6)/ml) and plasmid content (10 µg), the transfection efficiency of nuclear transfer apparatus default mode A-20 was higher than that of other modes (S-18, T-20). Its survival rate was up to 50.5% after 24 hours. Cell viability of liposome transfection reached to 88% after 24 hours, but the transfection efficiency was lower (< 1%). It is concluded that the nuclear transfer apparatus A-20 transfected L1210 can reach higher transfection efficiency up to 61.6%, which is significantly higher than that of lipofectamine transfection. The survival rate is up to 50.5% well meeting the needs of scientific research. Higher transfection efficiency is helpful for in-depth research about the morphology, functions and pathogenesis in leukemia model L1210, and provides more searching space for the treatment of leukemia diseases.
