Cell　surface　engineering　technologies　can　regulate　cell　function　and　behavior　by　modifying　the　cell　surface.　Previous　studies　have　mainly　focused　on　investigating　the　effects　of　cell　surface　engineering　reactions　and　materials　on　cell　activity.　However,　they　do　not　comprehensively　analyze　other　cellular　processes.　This　study　exploits　covalent　bonding,　hydrophobic　interactions,　and　electrostatic　interactions　to　modify　the　macromolecules　succinimide　ester-methoxy　polyethylene　glycol　(NHS-mPEG),　distearoyl　phosphoethanolamine-methoxy　polyethylene　glycol　(DSPE-mPEG),　and　poly-L-lysine　(PLL),　respectively,　on　the　cell　surface.　This　work　systematically　investigates　the　effects　of　the　three　surface　engineering　reactions　on　the　behavior　of　human　umbilical　vein　endothelial　cells　(HUVECs)　and　human　skin　fibroblasts,　including　viability,　growth,　proliferation,　cell　cycle,　adhesion,　and　migration.　The　results　reveals　that　the　PLL　modification　method　notably　affects　cell　viability　and　G2/M　arrest　and　has　a　short　modification　duration.　However,　the　DSPE-mPEG　and　NHS-mPEG　modification　methods　have　little　effect　on　cell　viability　and　proliferation　but　have　a　prolonged　modification　duration.　Moreover,　the　DSPE-mPEG　modification　method　highly　affects　cell　adherence.　Further,　the　NHS-mPEG　modification　method　can　significantly　improve　the　migration　ability　of　HUVECs　by　reducing　the　area　of　focal　adhesions.　The　findings　of　this　study　will　contribute　to　the　application　of　cell　surface　engineering　technology　in　the　biomedical　field.　©　2023　Wiley-VCH　GmbH.