The study of the interaction of a cationic polymer as PEI with phospholipids membranes is of special relevance for gene therapy because the PEI is a potential nonviral vector to transfer DNA in living cells. We used light scattering, zeta potential, and electron transmission microscopy to characterize the interaction between DMPG and DOPC liposomes with PEI as a function of the charge molar ratio, pH, temperature, initial size of the liposomes, and headgroup of the lipids. Unexpectedly, a double charge inversion and two different ranges of PEI-liposome concentrations where an aggregation occurs were found, when the proper pH and initial size of the liposomes were chosen. The interaction is analyzed in terms of the interaction potential proposed by Velegol and Thwar for colloidal particles with a nonuniform surface charge distribution. Results show a remarkable dependence of the stability on pH and the initial size of the liposomes, which explains the low reproducibility of the experiments if no special care is taken in preparing the samples. Comparatively small changes in the pH or in the liposomes size lead to a completely different stability behavior.