Optimization of gene transfer using cationic lipids in cell lines and primary human CD4+ and CD34+ hematopoietic cells

G. S. Harrison, Y. Wang, J. Tomczak, C. Hogan, E. J. Shpall, Tyler J Curiel, P. L. Felgner

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

Cationic lipids offer several advantages for gene delivery, both in vitro and in vivo. However, high-efficiency gene transfer has been demonstrated only for limited cell types. Here, we examine the level of expression of a luciferase reporter gene, delivered using cationic lipids, in both cell lines and primary human cells including peripheral blood mononuclear cells and CD34+-enriched hematopoietic cells. Variables shown to affect the efficiency of gene expression included the type of lipid, the amounts of DNA and lipid, the day of assay following transfection, the media used for lipid:DNA complex formation, the cell number, the promoter driving expression of the reporter gene and the physiological state of the cells (e.g., whether or not cells were differentiated). The maximal luciferase expression observed with the primary cells was one to two orders of magnitude lower than that seen in cell lines. Further studies, possibly involving altering the growth conditions for the cells, or using episomal vectors that will allow extrachromosomal maintenance of the DNA, are required to improve the level of transgene expression in the primary human cell types used here.

Original languageEnglish (US)
Pages (from-to)816-823
Number of pages8
JournalBioTechniques
Volume19
Issue number5
StatePublished - 1995
Externally publishedYes

Fingerprint

Gene transfer
Cells
Lipids
Cell Line
Genes
Luciferases
DNA
Reporter Genes
Gene expression
Assays
Blood
Transgenes
Transfection
Blood Cells
Cell Count
Maintenance
Gene Expression

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Clinical Biochemistry

Cite this

Harrison, G. S., Wang, Y., Tomczak, J., Hogan, C., Shpall, E. J., Curiel, T. J., & Felgner, P. L. (1995). Optimization of gene transfer using cationic lipids in cell lines and primary human CD4+ and CD34+ hematopoietic cells. BioTechniques, 19(5), 816-823.

Optimization of gene transfer using cationic lipids in cell lines and primary human CD4+ and CD34+ hematopoietic cells. / Harrison, G. S.; Wang, Y.; Tomczak, J.; Hogan, C.; Shpall, E. J.; Curiel, Tyler J; Felgner, P. L.

In: BioTechniques, Vol. 19, No. 5, 1995, p. 816-823.

Research output: Contribution to journalArticle

Harrison, GS, Wang, Y, Tomczak, J, Hogan, C, Shpall, EJ, Curiel, TJ & Felgner, PL 1995, 'Optimization of gene transfer using cationic lipids in cell lines and primary human CD4+ and CD34+ hematopoietic cells', BioTechniques, vol. 19, no. 5, pp. 816-823.
Harrison, G. S. ; Wang, Y. ; Tomczak, J. ; Hogan, C. ; Shpall, E. J. ; Curiel, Tyler J ; Felgner, P. L. / Optimization of gene transfer using cationic lipids in cell lines and primary human CD4+ and CD34+ hematopoietic cells. In: BioTechniques. 1995 ; Vol. 19, No. 5. pp. 816-823.
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