Journal of Jilin University Medicine Edition ›› 2016, Vol. 42 ›› Issue (03): 612-616.doi: 10.13481/j.1671-587x.20160338

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Optimization of liposome-mediated transfection of pEGFP-N1 and its applification

LI Fenge1, XIANG Guoyan2, KONG Fanli3, HAO Feng2, YANG Ziyan4, ZHENG Yan4, FANG Fang4   

  1. 1. Second Department of Neurology, Affiliated Hospital, Beihua University, Jilin 132013, China;
    2. Department of Pathophysiology, School of Basic Medical Sciences, Beihua University, Jilin 132013, China;
    3. Department of Biochemistry Laboratory, Jilin Medical College, Jilin 132013, China;
    4. Department of Pathogen, Jilin Medical College, Jilin 132013, China
  • Received:2016-01-17 Published:2016-06-17

Abstract:

Objective: To optimize the liposome-mediated transfection conditions based on transfecting the pEGFP-N1 into thyroid cells of Fischer rats,and to investigate its effectiveness on other target gene transfection. Methods: Based on the different ratios and quantities of plasmids and liposomes,the thyroid follicular epithelial cells of Fischer rats were transfected with pEGFP-N1.The expression of enhanced green fluorescence protein(EGFP) and transfection efficiency were observed under inverted fluorescence microscope and flow cytometry, respectively.Then the cell viability was detected by Trypan Blue stainning.Meanwhile the thyroid follicular epithelial cells of Fischer rats were transfected with pEGFP-Aquaporin2,pEGFP-Aquaporin4 and pEGFP-Aquaporin5 under the above conditions,and the optimized conditions of transfecion were obtained. Results: When the quantity of plasmid was certain,the transfection efficiency and cell viability were increased with the increasing of liposomes,but the transfection efficiency and cell viability were decreased when the ratio of plasmid to liposome was over 1:4.When the ratio of plasmid to liposome was certain and the quantities were increased gradully,the transfection effciency was increased,and the cell viability was decreased.With the ratio of plasmids to liposomes was 1:4 (3.2 μg:12.8 μL),the transfecion efficencies of pEGFP-N1,pEGFP-Aquaporin2,pEGFP-Aquaporin2, and pEGFP-Aquaporin2 reached the highest values,respectively. Conclusion: The optimized conditions of transfection pEGFP-N1 is the same as the other plasmids, which can be used as optimized conditions for efficient transfection of the other target gene.

Key words: liposome, pEGFP-N1, transfection, thyroid follicular epithelial cells

CLC Number: 

  • R361