RRC ID |
42241
|
著者 |
Liu Y, Uno H, Takatsuki H, Hirano M, Sakanishi A.
|
タイトル |
Interrelation between HeLa-S3 cell transfection and hemolysis in red blood cell suspension using pulsed ultrasound of various duty cycles.
|
ジャーナル |
Eur Biophys J
|
Abstract |
We have studied the in vitro transfection of a plasmid DNA with the lacZ gene to HeLa-S3 cells and hemolysis in a red blood cell (RBC) suspension under pulsed ultrasound with duty cycles gamma of 10, 20 and 30% using a digital sonifier at a frequency of 20 kHz and an intensity of 6.2 W/cm(2) on the surface of a horn tip. Cultured HeLa-S3 cells in suspension were exposed to pulsed ultrasound for an apparent exposure time t' from 0 to 60 s. HeLa-S3 viability decreased as a single exponential function of the total exposure time t = gammat' with a common time constant tau = 3.8 s for three duty cycles. Transfection was evaluated by counting the number of beta-galactosidase(beta-Gal)-positive cells relative to the total number of cells. Pulsed ultrasound provided an enhanced transfer of the beta-Gal plasmid to HeLa-S3 cells, 3.4-fold as compared with that in the case of the control. The optimal transfection efficiencies were 0.75, 0.80 and 0.74% near t = tau with gamma = 10, 20 and 30%, respectively. The number ratio of beta-Gal-positive cells to the surviving cells after exposure increased with t' according to a modified logistic equation. The degree of hemolysis also increased exponentially with t' at a time constant tau' = tau(0)/gamma for the RBC suspension in physiological saline at a hematocrit concentration of 0.5% with tau(0) = 0.9 s. Thus the total exposure time for the optimal transfection efficiency was tau, that is, nearly four times of tau(0). Hemolysis in the RBC suspension may be a useful model for determining optimal transfection by pulsed ultrasound of various duty cycles.
|
巻・号 |
34(2)
|
ページ |
163-9
|
公開日 |
2005-3-1
|
DOI |
10.1007/s00249-004-0439-7
|
PMID |
15480621
|
MeSH |
Animals
Cell Membrane Permeability / radiation effects
Cell Survival / radiation effects
Cells, Cultured
Dose-Response Relationship, Radiation
Erythrocytes / drug effects
Erythrocytes / physiology*
Genetic Therapy / methods
HeLa Cells
Hemolysis / physiology*
Hemolysis / radiation effects
Humans
Radiation Dosage
Swine
Transfection / methods*
Ultrasonics*
|
IF |
2.094
|
引用数 |
6
|
WOS 分野
|
BIOPHYSICS
|
リソース情報 |
ヒト・動物細胞 |
HeLa.S3(RCB0191) |