Optimization of the transfection of human THP-1 macrophages by application of Nunc UpCell technology

Anal Biochem. 2015 Jun 15:479:40-2. doi: 10.1016/j.ab.2014.12.023. Epub 2015 Feb 4.

Abstract

We have established an electroporation protocol for transfection of premature adherent human THP-1 macrophages using Lonza Nucleofector technology. For efficient electroporation, detachment of adherent cells is necessary. We tested the Nunc UpCell product line of Thermo Fisher Scientific, which achieves detachment by a change of ambient temperature, as an alternative to enzymatic detachment. Here we present data verifying proper cell morphology and vitality and high transfection efficiency for macrophages cultured on UpCell plates. Appropriate macrophage behavior was confirmed by measuring markers of macrophage differentiation and polarization by reverse transcription quantitative polymerase chain reaction (RT-qPCR). In conclusion, Nunc UpCell materials are a viable alternative to enzymatic detachment.

Keywords: Detachment; THP-1 macrophages; Transfection; siRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Adhesion
  • Cell Differentiation
  • Cell Line
  • Cell Survival
  • Electroporation / methods*
  • Humans
  • Macrophages / cytology
  • Macrophages / metabolism*
  • RNA Interference*
  • RNA, Small Interfering / administration & dosage*
  • RNA, Small Interfering / genetics
  • Transfection / methods*

Substances

  • RNA, Small Interfering