title: A rapid and efficient polyethylenimine-based transfection method to prepare lentiviral or retroviral vectors: useful for making iPS cells and transduction of primary cells.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Stem Cell P2 Laboratory, Shantou University Medical College, Shantou, 515041, People's Republic of China.|The Center for Reproductive Medicine, Shantou University Medical College, Shantou, 515041, People's Republic of China.|Guangdong Provincial Key Laboratory of Infectious Diseases and Molecular Immunopathology, Shantou University Medical College, Shantou, 515041, People's Republic of China.
authors.forename: Shaozhe
authors.initials: S
authors.lastname: Yang
authors.affiliation: Stem Cell P2 Laboratory, Shantou University Medical College, Shantou, 515041, People's Republic of China.|The Center for Reproductive Medicine, Shantou University Medical College, Shantou, 515041, People's Republic of China.|Guangdong Provincial Key Laboratory of Infectious Diseases and Molecular Immunopathology, Shantou University Medical College, Shantou, 515041, People's Republic of China.
authors.forename: Haijun
authors.initials: H
authors.lastname: Shi
authors.affiliation: Stem Cell P2 Laboratory, Shantou University Medical College, Shantou, 515041, People's Republic of China.|The Center for Reproductive Medicine, Shantou University Medical College, Shantou, 515041, People's Republic of China.|Guangdong Provincial Key Laboratory of Infectious Diseases and Molecular Immunopathology, Shantou University Medical College, Shantou, 515041, People's Republic of China.
authors.forename: Xinran
authors.initials: X
authors.lastname: Chu
authors.affiliation: Stem Cell P2 Laboratory, Shantou University Medical College, Shantou, 515041, People's Republic of China.|The Center for Reproductive Medicine, Shantou University Medical College, Shantou, 515041, People's Republic of China.|Guangdong Provincial Key Laboratory of Infectious Diseases and Molecular Immunopathology, Shantou University Medical College, Shantou, 515041, People's Republic of China.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Stem Cell P2 Laboratory, Shantou University Medical College, Shantou, 515041, People's Republic of China. Pingnan_sun@yahoo.com.|The Center for Reproductive Medicine, Shantou University Medical College, Shantou, 515041, People's Republic of China. Pingnan_sun@yahoo.com.|Guangdong Provincial Key Laboratory of Infectious Diseases and Molecular Immunopathology, Shantou University Medical College, Shantou, 515041, People's Republic of China. Pingnan_sun@yahoo.com.
authors.forename: Pingnan
authors.initials: P
authors.lastname: Sun
mesh_terms: D000818:Animals; D002460:Cell Line; D005822:Genetic Vectors / Q000737:chemistry*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D011094:Polyethyleneimine / Q000737:chemistry*; D012190:Retroviridae / Q000235:genetics*; D014161:Transduction, Genetic / Q000379:methods*; D014162:Transfection / Q000379:methods*
chemical_list: D011094:Polyethyleneimine
keywords: Lentivirus; Polyethylenimine; Retrovirus; Transduction; Transfection