Oligonucleotide uptake in cultured keratinocytes: Influence of confluence, cationic liposomes, and keratinocyte cell type

Paul J. White, Rhys D. Fogarty, Sandra C. McKean, Daryl J. Venables, George A. Werther, Christopher J. Wraight

Research output: Contribution to journalArticlepeer-review

28 Citations (Scopus)

Abstract

The success of anti-sense strategies has been limited, at least in part, by the poor uptake of these agents into the target cells. In keratinocytes, there is conflicting evidence as to the amount and location of oligonucleotide uptake into these cells, with variable proportions of cells reported to take up oligodeoxynucleotide, and also cytoplasmic and nuclear localization reported. In this study, the uptake of oligodeoxynucleotides in cultured normal human keratinocytes and the HaCaT cell line was quantitated in the presence of various lipids designed to enhance uptake and in varying culture conditions. About 12% of cells in a confluent normal human keratinocyte culture showed nuclear uptake, with a small and variable proportion showing cytoplasmic localization after 24 h incubation with 1 μM oligodeoxynucleotide. Uptake of oligodeoxynucleotide was found to be increased by liposome encapsulation (to a maximum of 28.1% ± 2.1% of cells), low confluence (39.5% ± 2.5%), and further increased by a combination of the two conditions (55.4% ± 4.3%). HaCaT cell populations showed sparse but consistent uptake of oligodeoxynucleotide, with about 1% of cells showing nuclear localization in the presence of 1 μM oligodeoxynucleotide, increasing to 13.5% ± 4.9% in the presence of cationic lipid (Tfx-50) in low confluence HaCaT monolayers. We conclude that normal keratinocytes exhibit reliable, substantial uptake of oligonucleotides in conditions controlled for confluence and aided by liposome encapsulation.

Original languageEnglish
Pages (from-to)699-705
Number of pages7
JournalJournal of Investigative Dermatology
Volume112
Issue number5
DOIs
Publication statusPublished or Issued - 1999

Keywords

  • Anti-sense
  • Delivery
  • Propyne

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Dermatology
  • Cell Biology

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