Introduction of macromolecules into mammalian cells by cell fusion.
Proteins with molecular weights of up to 500K can be enclosed in erythrocyte ghosts by exposing the ghosts to hypotonic solution containing these proteins. The proteins can then be introduced into recipient cells by fusing the ghosts with the cells using HVJ, PEG, or influenza virus. Some applications of this method are described. By an improved method, 15 kbp DNA and IgM (900 kDa) can be entrapped in erythrocyte membranes and these are then treated with liposomes containing gangliosides and HVJ. These treated membranes containing large macromolecules fuse with almost 100% of the recipient cells used. Naked liposomes infrequently fuse with cultured cells, so introduction of their contents into cells is very inefficient. However, liposomes constituted from lipid and glycoproteins (HN and F) of HVJ (Sendai virus), by removing a nonionic detergent, fuse with cells about 200 times more efficiently than naked liposomes. Naked liposomes can fuse with specific cells, such as cells infected with subacute sclerosing panencephalitis virus or with human immunodeficiency virus. Plasmid DNA and mRNA of up to about 40 kbp can be entrapped efficiently in liposomes associated with gangliosides formed by reverse-phase evaporation, and then reacted with HVJ. The contents of the resulting liposomes with HVJ can be introduced efficiently into cultured cells in a suspended or plated state, and nearly all the cells then express the gene transiently. This procedure is also effective for obtaining stable transformants of many kinds of cultured cells.
Uchida
T
T
Institute for Molecular and Cellular Biology, Osaka University, Japan.
D002459:Cell Fusion; D004910:Erythrocyte Membrane; D006801:Humans; D066298:In Vitro Techniques; D008081:Liposomes; D046911:Macromolecular Substances; D010222:Parainfluenza Virus 1, Human
D008081:Liposomes; D046911:Macromolecular Substances