Phosphate-buffered saline-based nucleofection of primary endothelial cells

Jinjoo Kang, Swapnika Ramu, Sunju Lee, Berenice Aguilar, Sathish Kumar Ganesan, Jaehyuk Yoo, Vijay K. Kalra, Chester J. Koh, Young Kwon Hong

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

Although various nonviral transfection methods are available, cell toxicity, low transfection efficiency, and high cost remain hurdles for in vitro gene delivery in cultured primary endothelial cells. Recently, unprecedented transfection efficiency for primary endothelial cells has been achieved due to the newly developed nucleofection technology that uses a combination of novel electroporation condition and specific buffer components that stabilize the cells in the electrical field. Despite superior transfection efficiency and cell viability, high cost of the technology has discouraged cardiovascular researchers from liberally adopting this new technology. Here we report that a phosphate-buffered saline (PBS)-based nucleofection method can be used for efficient gene delivery into primary endothelial cells and other types of cells. Comparative analyses of transfection efficiency and cell viability for primary arterial, venous, microvascular, and lymphatic endothelial cells were performed using PBS. Compared with the commercial buffers, PBS can support equally remarkable nucleofection efficiency to both primary and nonprimary cells. Moreover, PBS-mediated nucleofection of small interfering RNA (siRNA) showed more than 90% knockdown of the expression of target genes in primary endothelial cells. We demonstrate that PBS can be an unprecedented economical alternative to the high-cost buffers or nucleofection of various primary and nonprimary cells.

Original languageEnglish (US)
Pages (from-to)251-255
Number of pages5
JournalAnalytical Biochemistry
Volume386
Issue number2
DOIs
StatePublished - Mar 15 2009

Keywords

  • Electroporation
  • Nucleofection
  • Phosphate-buffered saline
  • Primary endothelial cells

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Cell Biology

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