TRANSFECTION OF MYOBLASTS IN PRIMARY CULTURE WITH ISOMERIC CATIONIC CHOLESTEROL DERIVATIVES

Transfection of satellite cells from dog muscle (myoblasts) in primary culture has been optimized with respect to the position of the choles teryl moiety along the polyamine chain of spermidine or spermine. Sper midine or spermine were derivatized with cholesterylchloroformate givi ng rise to three isomers in the case of spermidine and two isomers for spermine that were separated by reversed-phase high-performance liqui d chromatography (rp-HPLC). The position of the cholesteryl moiety was assigned by C-13-NMR and coelution with synthetic isomers of defined structure. The isomeric cationic lipids were evaluated for their trans fection activity in myoblasts from dog muscle and a human lung epithel ial cell line (A549) using plasmid DNA expressing the luciferase repor ter gene. The results showed that the position of the cholesteryl moi ety is of critical importance for efficient transfection of myoblasts in primary culture with isomers having a derivatized secondary amine b eing significantly more effective than those with a derivatized primar y amine. On the contrary, differences in the A549 cell line were less pronounced and did not follow the same pattern, The results show that slight structural differences between cationic lipids lead to signific antly different transfection efficiencies for myoblasts in primary cul ture. This may also represent an advantage in view of cell or organ ta rgeting. (C) 1997 Academic Press.