Efficient introduction of macromolecules and oligonucleotides into brain capillary endothelial cells using HVJ-liposomes.
In this study, we examined the feasibility of introducing macromolecules into cultured mouse brain capillary endothelial cells (MBEC4 cells) by utilizing the hemagglutating virus of Japan (HVJ)-liposomes with fusogenic activity. We used fluorescein isothiocyanate dextran (FITC-Dextran) and FITC-labeled oligodeoxynucleotide (FITC-ODN) as models of a macromolecule and an ODN, respectively. Intracellular fluorescence appeared rapidly after the exposure of MBEC4 cells to FITC-Dextran-containing HVJ-liposomes, and remained detectable for at least 3 days. Only a control level of intracellular fluorescence was seen after treatment with FITC-Dextran alone, FITC-Dextran with empty HVJ-liposomes or FITC-Dextran-containing liposomes without fusogenic activity. In the early phase after administration (0-30 min), the introduction of FITC-Dextran into MBEC4 cells by the HVJ-liposome method resulted in a rapid and time-dependent increase of intracellular fluorescence intensity. Moreover, FITC-ODN was also introduced into MBEC4 cells by the HVJ-liposome method, although FITC-ODN alone was not introduced. These results indicate that the HVJ-liposome method is useful for the efficient introduction of macromolecules, including ODN, into brain capillary endothelial cells.
Matsuo
H
H
Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Okamura
T
T
Chen
J
J
Takanaga
H
H
Ohtani
H
H
Kaneda
Y
Y
Naito
M
M
Tsuruo
T
T
Sawada
Y
Y
D000818:Animals; D001693:Biological Transport, Active; D001812:Blood-Brain Barrier / Q000502:physiology; D001921:Brain / Q000098:blood supply*; D002460:Cell Line; D003911:Dextrans / Q000008:administration & dosage* / Q000493:pharmacokinetics; D016503:Drug Delivery Systems; D004730:Endothelium, Vascular / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate / Q000008:administration & dosage* / Q000031:analogs & derivatives* / Q000493:pharmacokinetics; D008081:Liposomes / Q000008:administration & dosage*; D046911:Macromolecular Substances; D008561:Membrane Fusion / Q000502:physiology; D051379:Mice; D008856:Microscopy, Fluorescence; D009841:Oligonucleotides / Q000008:administration & dosage* / Q000493:pharmacokinetics; D018108:Respirovirus; D013379:Substrate Specificity
D003911:Dextrans; D008081:Liposomes; D046911:Macromolecular Substances; D009841:Oligonucleotides; C015219:fluorescein isothiocyanate dextran; D016650:Fluorescein-5-isothiocyanate