Evaluation of peptide-mediated transduction in human CD34+ cells

Hum Gene Ther. 2004 Apr;15(4):415-9. doi: 10.1089/104303404322959560.

Abstract

Protein transduction domains (PTDs) have been used increasingly to deliver biologically active agents to a variety of cell types in vitro and in vivo. To define the most effective PTDs for transducing hematopoietic cells, we have screened a panel of PTD peptides in human CD34(+) cells for delivery of a 60-kd marker protein and assessed its impact on phenotypic maintenence in vitro. Compared to the HIV-TAT peptide, most peptide complexes displayed high efficiency in transducing the CD34(+) cells, except for those based on shorter peptides (4R, 4K, and 5RQ). In particular, the arginine homopolymers including 8R, 10R, and 12R, were internalized by the cells to a greater extent than the other PTDs. Transduction was significantly potentiated by preincubation of cells with dextran sulfate. Importantly, colony forming ability and CD34(+) CD38(-) primitive phenotype were not significantly altered in the presence of these peptides during a short-term liquid culture. Together, these data suggest the potential usefulness of arginine homopolymers in hematopoietic stem and progenitor cell manipulations.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antigens, CD34 / metabolism*
  • Colony-Forming Units Assay
  • Dextran Sulfate / pharmacology*
  • Fetal Blood
  • Humans
  • Peptide Fragments / chemistry
  • Peptide Fragments / metabolism*
  • Protein Structure, Tertiary
  • Transduction, Genetic*
  • Transfection

Substances

  • Antigens, CD34
  • Peptide Fragments
  • Dextran Sulfate