We have designed α-helical peptides de novo that can induce aggregation of various kinds of cells by focusing on physicochemical properties such as hydrophobicity, net charges, and amphipathicity. It is shown that peptide hydrophobicity is the key factor to determine capabilities for cell aggregation while peptide net charges contribute to nonspecific electrostatic interactions with cells. On the other hand, amphipathic peptides tend to exhibit cytotoxicity such as antimicrobial activity and hemolysis, which are competitive with cell-aggregation capabilities. Different from the cases of living cells, aggregation of artificial anionic liposomes appears to be mainly determined by electrostatic interactions. This discrepancy might be due to the complex structure of surfaces of cell membranes consisting of macromolecular chains such as peptidoglycans, polysaccharides, or glycocalyx, which coexist with lipid bilayers. Our design strategy would pave the way to design peptides that lead aggregation of living cells without cytotoxicity.