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Comparison of Different Cell Transfection by Cationic Polymer

Author: ZhangLi
Tutor: XuHong；XiongWei
School: Jilin University
Course: Internal Medicine
Keywords: gene transfection polyethylenimine (PEI) Lipofectamine2000 murine skeletal muscle C2C12 human umbilical vein endothelial cells HUVEC
CLC: R450
Type: Master's thesis
Year: 2011
Downloads: 75
Quote: 1
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Abstract

Background Gene therapy refers to the foreign genes into the target cell interior through restoring or adding gene expression of genes to correct people’s the disorder of own structure or function,which prevent the progress of disease and kill disease cells or inhibit replication of exogenous pathogens genetic material to achieve the purpose of treatment.The gene therapy has three important links, namely the goal gene,the transgene vector and the target cell.The gene carrier mainly divides into the viral carrier system and the non-viral carrier system.The polyethylenamine (PEI)have attracted more and more attention because they have many advantages in terms of low toxicity,lack of specific immune response,ease of large-scale production and big load.PEI already succeeded uses in vivo and in vitro gene transfer at present.Experiments have demonstrated that gene transfer efficiency of the PEI-PBLG after modification of PEI is better than polyethylenimine (PEI), simultaneously the cytotoxicity reduces obviously.Similarly the choice different target cell is also playing the vital role to the gene therapy, therefore the choice correct cell subline carries on vitro experiment to be helpful to further enhances the gene transfer efficiency.This experiment makes comparison and observation by Lipofectamine2000,PEI and the PEI-PBLG transfection efficiency and cytotoxicity of cationic liposome in different cells,which provides the reference for the further in vivo experiment.Objective:This research transfects different cell human cervical carcinoma cell Hela, murine skeletal muscle C2C12 and human umbilical vein endothelial cells HUVEC through the contrast different material in vitro and examines gene transfection efficiency and toxic effect of material to cells.Method:With cationic liposome Lipofectamine2000,polycation polyethylenimine (PEI) and the modified materials (PEI-PBLG) as vector, the transfection efficiency of the materialto different cells (Hela,C2C12 and HUVEC) were studied and evaluated through plasmid transfection test. The latter two materials to three kind of cell’s toxic effects were examined through the MTT.Result:EGFP in three kind of cell subline by different types of vector-mediated gene transfection was found,and their expression increased 24 hours after transfection with a summit at hour 48.The highest transfection efficiency cells Hela, followed by C2C12, HUVCE. The efficiency of three materials were Lipofectamine2000> PEI-PBLG> PEI. Lipofectamine2000 to three cells in which the highest transfection efficiency were 63%,18%,17%.It is same that toxic effect of PEI and PEI-PBLG to three kind of cells,but the toxicity of PEI-PBLG higher than PEI.Conclusion:Experiments by in vitro show that the transfection efficiency of epithelial cells (Hela) higher than murine skeletal muscle (C2C12) and human vascular endothelial cells (HUVEC).The C2C12 cell and the HUVEC cell trans gene efficiency is similar and low.Two materials are consistent to three kind of cell’s toxic effects, which is the PEI-PBLG toxicity is smaller than PEI.It Provides a further refers for method of choice in vivo testing.

Comparison of Different Cell Transfection by Cationic Polymer

Abstract

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