S. Novakovic et al., Transfection of mammalian cells by the methods of receptor mediated gene transfer and particle bombardment, J EXP CL C, 18(4), 1999, pp. 531-536
Citations number
15
Categorie Soggetti
Oncology
Journal title
JOURNAL OF EXPERIMENTAL & CLINICAL CANCER RESEARCH
The efficacy of currently developed methods for gene transfer into mammalia
n cells depends primarily on the transfection technique, and also on the ty
pe of targeted cells. Considering the importance of gene transfer in the cr
eation of gene therapies, our study was aimed at the assessment of transfec
tion capacity of receptor mediated gene transfer method (RMGT), and method
of particle bombardment (helios gene gun system - HGG) in different normal
and malignant mammalian cells ex vivo. In addition, the HGG was also assess
ed for its ability to transfect tumor cells of subcutaneous (s.c.) tumors i
n C57B1/6 mice in vivo. Using RMGT an average ex vivo transfection rate of
35.7%; and 20.4% was achieved in malignant melanoma B-16, and human breast
adenocarcinoma MCF7, respectively. However, in normal fibroblast L929 cells
the transfection by RMGT succeeded only in 2.1% of the cells. On the other
hand, the transfection efficacy of HGG was comparable in both malignant ce
ll lines resulting in an average gene transfer to 9.6% of B-16 and 10.5% of
MCF7 cells, while only 3.9% of normal fibroblasts were successfully transf
ected. Application of HGG for an in vivo gene transfer into s.c, B-16 melan
oma tumors in C57B1/6 mice resulted in a successful but limited transfectio
n of the epithelium as well as of the superficially sited tumor cells. Taki
ng into consideration both methods, RMGT is more appropriate for ex vivo tr
ansfection of cells, under the condition that target cells express a specif
ic receptor for the molecule attached to the carrier. On the other hand, HG
G is not complicated to use, no requirements for specific structures on tar
get cells are necessary (potentially usable in different cells), and it has
applicability in direct in vivo transfection processes.