Differential scanning calorimetry was used to examine the lipid exchange between model lipid systems, including vesicles of the cationic lipoids ethyldimyristoylphosphatidylcholine (EDMPC), ethyldipalmitoylphosphatidylcholine (EDPPC) or their complexes with DNA (lipoplexes), and the zwitterionic lipids (DMPC, DPPC). The changes of the lipid phase transition parameters (temperature, enthalpy, and cooperativity) upon consecutive temperature scans was used as an indication of lipid mixing between aggregates. A selective lipid transfer of the shorter-chain cationic lipoid EDMPC into the longer-chain aggregates was inferred. In contrast, transfer was hindered when EDMPC (but not EDPPC) was bound to DNA in the lipoplexes. These data support a simple molecular lipid exchange mechanism, but not lipid bilayer fusion. Exchange via lipid monomers is considerably more facile for the cationic ethylphosphatidylcholines than for zwitterionic phosphatidylcholines, presumably due to the higher monomer solubility of the charged lipids. With the cationic liposomes, lipid transfer was strongly promoted by the presence of serum in the dispersing medium. Serum proteins are presumed to be responsible for the accelerated transfer, since the effect was strongly reduced upon heating the serum to 80 degrees C. The effect of serum indicates that even though much lipoplex lipid is inaccessible due to the multilayered structure, the barrier due to buried lipid can be easily overcome. Serum did not noticeably promote the lipid exchange of zwitterionic liposomes. The phenomenon is of potential importance for the application of cationic liposomes to nonviral gene delivery, which often involves the presence of serum in vitro, and necessarily involves serum contact in vivo.