Evaluation of peptide-mediated transduction in human CD34+ cells.
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.
Shen
Hongmei
H
University of Pittsburgh Cancer Institute, Department of Radiation Oncology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261.
Mai
Jeffrey C
JC
Qiu
Lu
L
Cao
Shaonan
S
Robbins
Paul D
PD
Cheng
Tao
T
D018952:Antigens, CD34 / Q000378:metabolism*; D003114:Colony-Forming Units Assay; D016264:Dextran Sulfate / Q000494:pharmacology*; D005312:Fetal Blood; D006801:Humans; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D014161:Transduction, Genetic; D014162:Transfection
D018952:Antigens, CD34; D010446:Peptide Fragments; D016264:Dextran Sulfate