Quantitative study of electroporation-mediated molecular uptake and cell viability.
Electroporation's use for laboratory transfection and clinical chemotherapy is limited by an incomplete understanding of the effects of electroporation parameters on molecular uptake and cell viability. To address this need, uptake of calcein and viability of DU 145 prostate cancer cells were quantified using flow cytometry for more than 200 different combinations of experimental conditions. The experimental parameters included field strength (0.1-3.3 kV/cm), pulse length (0.05-20 ms), number of pulses (1-10), calcein concentration (10-100 microM), and cell concentration (0.6-23% by volume). These data indicate that neither electrical charge nor energy was a good predictor of electroporation's effects. Instead, both uptake and viability showed a complex dependence on field strength, pulse length, and number of pulses. The effect of cell concentration was explained quantitatively by electric field perturbations caused by neighboring cells. Uptake was shown to vary linearly with external calcein concentration. This large quantitative data set may be used to optimize electroporation protocols, test theoretical models, and guide mechanistic interpretations.
Canatella
P J
PJ
School of Chemical Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
Karr
J F
JF
Petros
J A
JA
Prausnitz
M R
MR
D001693:Biological Transport, Active; D055592:Biophysical Phenomena; D001703:Biophysics; D002470:Cell Survival; D018274:Electroporation / Q000379:methods*; D005434:Flow Cytometry; D005452:Fluoresceins / Q000378:metabolism; D006801:Humans; D008297:Male; D008954:Models, Biological; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology; D014407:Tumor Cells, Cultured
D005452:Fluoresceins; C007740:fluorexon