Gene introduction into granulocyte-macrophage progenitor cells by electroporation: the relationship between introduction efficiency and the proportion of cells in S-phase.
Our previous study demonstrated the positive relationship between the gene introduction rate into hematopoietic cell lines by electroporation and the percentage of cells in S-phase. In the present study, granulocyte-macrophage progenitor cells (CFU-C) rich marrow cell fraction were cultured in suspension with IL-3, GM-CSF and G-CSF for 4 days. The number of CFU-C were increased three times after the culture, and 3H-thymidine suicide tests of cultured cells demonstrated that the proportion of CFU-C in S-phase was increased by two to four times. The efficiency of gene transfer into CFU-C with the plasmid pMoZtk (containing the beta-galactosidase gene) by electroporation was nearly doubled by culturing marrow cells with these growth factors. These findings confirm that the introduction rate of the gene into CFU-C by electroporation is more efficient in cell populations with a higher percentage of CFU-C in S-phase.
Takahashi
M
M
First Department of Internal Medicine, Niigata University School of Medicine, Japan.
Furukawa
T
T
Tanaka
I
I
Nikkuni
K
K
Aoki
A
A
Kishi
K
K
Koike
T
T
Moriyama
Y
Y
Shibata
A
A
D002452:Cell Count; D003114:Colony-Forming Units Assay; D005821:Genetic Techniques; D006098:Granulocytes / Q000201:enzymology / Q000473:pathology*; D006412:Hematopoietic Stem Cells / Q000201:enzymology / Q000473:pathology*; D006801:Humans; D008264:Macrophages / Q000201:enzymology / Q000473:pathology*; D016196:S Phase; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
D001616:beta-Galactosidase