A rapid and efficient polyethylenimine-based transfection method to prepare lentiviral or retroviral vectors: useful for making iPS cells and transduction of primary cells

Objectives To improve the efficiency, reproducibility and consistency of the PEI-based transfection method that is often used in preparation of recombinant lentiviral or retroviral vectors. Results The contributions to transfection efficiency of multi-factors including concentration of PEI or DNA, d...

Full description

Saved in:
Bibliographic Details
Published in:Biotechnology letters 2016-09, Vol.38 (9), p.1631-1641
Main Authors: Yang, Shaozhe, Shi, Haijun, Chu, Xinran, Zhou, Xiaoling, Sun, Pingnan
Format: Article
Language:English
Subjects:
Animals
Applied Microbiology
Biochemistry
Biomedical and Life Sciences
Biotechnology
Cell Line
Cellular biology
Consistency
Culture
Deoxyribonucleic acid
DNA
Genetic Vectors - chemistry
Humans
Lentivirus - genetics
Life Sciences
Mathematical analysis
Microbiology
Original Research Paper
Polyetherimides
Polyethyleneimine - chemistry
Recombinant
Retroviridae - genetics
Signal transduction
Transduction, Genetic - methods
Transfection - methods
Vectors (mathematics)
Viruses
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Objectives To improve the efficiency, reproducibility and consistency of the PEI-based transfection method that is often used in preparation of recombinant lentiviral or retroviral vectors. Results The contributions to transfection efficiency of multi-factors including concentration of PEI or DNA, dilution buffer for PEI/DNA, manner to prepare PEI/DNA complexes, influence of serum, incubation time for PEI/DNA complexes, and transfection time were studied. Gentle mixing during the preparation of PEI/DNA transfection complexes is critical for a high transfection efficiency. PEI could be stored at room temperature or 4 °C, and most importantly, multigelation should be avoided. The transfection efficiency of the PEI-based new method in different types of cells, such as 293T, Cos-7, HeLa, HepG2, Hep3B, Huh7 and L02, was also higher than that of the previous method. After optimization, the titer of our lentiviral system or retroviral system produced by PEI-based new method was about 10- or 3-times greater than that produced by PEI-based previous method, respectively. Conclusion We provide a rapid and efficient PEI-based method for preparation of recombinant lentiviral or retroviral vectors which is useful for making iPS cells as well as transduction of primary cell cultures.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-016-2123-2