Increased liposome-mediated gene transfer into haematopoietic cells grown in adhesion to stromal or fibroblast cell line monolayers.
We investigated transfection rates of CD34+ haematopoietic progenitor cells (HPC) or haematopoietic cell lines (TF-1, KG1a and K562) using the LacZ gene as a reporter and cationic liposomes. The transfection efficiency of CD34+ haematopoietic progenitor cells (HPC) or TF-1, KG1a and K562 grown in suspension is very low (average percentage of 0.013 for HPC and 0.03 for cell lines). Adhesion of HPC or cell lines to plates by immunological or physical methods significantly enhances transfection efficiency; however, the percentage of transfected cells still remained low. We found that adhesion of TF-1, KG1a and K562 HC to MS-5 stroma cells or NIH-3T3 fibroblast cells increased transfection efficiency. Under these conditions transfection is achieved in 11.2-25% (mean 18.30%) for the cell lines and 13.6% (range 8.2-24.2%) for CD34+ HPC. These results indicate that liposome-mediated transfection of HC is significantly increased when cells are grown in adherence to stroma or fibroblast monolayers.
Marit
G
G
Laboratoire de Greffe de Moelle, UMR CNRS 5540, Université Victor Segalen Bordeaux 2, France. Gerald.Marit@umr5540.u-bordeaux2.fr
Cao
Y
Y
Froussard
P
P
Ripoche
J
J
Dupouy
M
M
Elandaloussi
A
A
Lacombe
F
F
Mahon
F X
FX
Keller
H
H
Pla
M
M
Reiffers
J
J
Theze
J
J
D016475:3T3 Cells; D000818:Animals; D002411:Cation Exchange Resins; D002448:Cell Adhesion; D002460:Cell Line; D002478:Cells, Cultured; D018920:Coculture Techniques; D004337:Drug Carriers; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D017930:Genes, Reporter; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000502:physiology*; D006801:Humans; D020014:K562 Cells; D007700:Kinetics; D008055:Lipids; D008081:Liposomes; D051379:Mice; D017154:Stromal Cells / Q000166:cytology* / Q000502:physiology; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000235:genetics*
D002411:Cation Exchange Resins; D004337:Drug Carriers; D008055:Lipids; C086724:Lipofectamine; D008081:Liposomes; D001616:beta-Galactosidase