In vitro gene transfer to mammalian cells by the use of laser-induced stress waves: effects of stress wave parameters, ambient temperature, and cell type.
Laser-mediated gene transfection has received much attention as a new method for targeted gene therapy because of the high spatial controllability of laser energy. We previously demonstrated both in vivo and in vitro that plasmid DNA can be transfected by applying nanosecond pulsed laser-induced stress waves (LISWs). In the present study, we investigated the dependence of transfection efficiency on the laser irradiation conditions and hence stress wave conditions in vitro. We measured characteristics of LISWs used for gene transfection. For NIH 3T3 cells, transfection efficiency was evaluated as functions of laser fluence and number of pulses. The effect of ambient temperature was also investigated, and it was found that change in ambient temperature in a specific range resulted in drastic change in transfection efficiency for NIH 3T3 cells. Gene transfection of different types of cell lines were also demonstrated, where cellular heating increased transfection efficiency for nonmalignant cells, while heating decreased transfection efficiency for malignant cells.
Terakawa
Mitsuhiro
M
Keio University, Department of Electronics and Electrical Engineering, 3-14-1, Hiyoshi, Yokohama, Kanagawa 223-8522, Japan. tera@obara.elec.keio.ac.jp
Sato
Shunichi
S
Ashida
Hiroshi
H
Aizawa
Kazuya
K
Uenoyama
Maki
M
Masaki
Yoshinori
Y
Obara
Minoru
M
D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism* / Q000528:radiation effects*; D002463:Cell Membrane Permeability / Q000528:radiation effects; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D004307:Dose-Response Relationship, Radiation; D005347:Fibroblasts / Q000378:metabolism* / Q000528:radiation effects; D006367:HeLa Cells; D006801:Humans; D007834:Lasers; D051379:Mice; D041681:NIH 3T3 Cells; D010957:Plasmids / Q000235:genetics* / Q000493:pharmacokinetics*; D011829:Radiation Dosage; D013045:Species Specificity; D013314:Stress, Mechanical; D013696:Temperature; D014162:Transfection / Q000379:methods*