Optimal lentivirus production and cell culture conditions necessary to successfully transduce primary human bronchial epithelial cells

Nathalie Baumlin-Schmid, Matthias A Salathe, Nevis L. Fregien

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

In vitro culture of primary human bronchial epithelial (HBE) cells using air-liquid interface conditions provides a useful model to study the processes of airway cell differentiation and function. In the past few years, the use of lentiviral vectors for transgene delivery became common practice. While there are reports of transduction sof fully differentiated airway epithelial cells with certain non-HIV pseudo-typed lentiviruses, the overall transduction efficiency is usually less than 15%. The protocol presented here provides a reliable and efficient method to produce lentiviruses and to transduce primary human bronchial epithelial cells. Using undifferentiated bronchial epithelial cells, transduction in bronchial epithelial growth media, while the cells attach, with a multiplicity of infection factor of 4 provides efficiencies close to 100%. This protocol describes, step-by-step, the preparation and concentration of high-titer lentiviral vectors and the transduction process. It discusses the experiments that determined the optimal culture conditions to achieve highly efficient transductions of primary human bronchial epithelial cells.

Original languageEnglish (US)
Article numbere54176
JournalJournal of Visualized Experiments
Volume2016
Issue number113
DOIs
StatePublished - Jul 22 2016

Keywords

  • Airliquid interface medium
  • Bronchial epithelial growth medium
  • Issue 113
  • Lentiviral vector
  • Microbiology
  • Multiplicity of infection
  • Primary human bronchial epithelial cells
  • Transduction

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

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