Multiresponsive　and　biocompatible　self-healing　ϵ-PL/A-Pul/BPEI　hydrogels　were　prepared　in　aqueous　solution　by　Schiff　base　reaction　with　aldehyded　pullulan　(A-Pul),　ϵ-poly-l-lysine　(ϵ-PL)　and　branched　polyethyleneimine　(BPEI)　as　materials.　The　imine　bonds　were　rapidly　cross-linked　into　a　hydrogel　network　within　80　s.　Scanning　electron　microscopy　images　showed　that　the　hydrogels　exhibited　a　cross-linked　structure　with　the　average　pore　size　from　58　to　82　μm.　Rheology　tests　indicated　that　the　hydrogels　maintained　good　mechanical　properties.　Water　contact　angles　and　swelling　studies　suggested　that　the　hydrogels　could　swell　in　water,　with　a　max　swell　ratio　of　1559%,　and　pH　and　temperature　had　an　influence　on　the　equilibrium　swelling　ratio.　The　hydrogels　could　be　injected　either　before　or　after　gelation,　and　they　displayed　a　self-healing　process　in　ddH2O　at　room　temperature　based　on　the　dynamic　uncoupling　and　recoupling　of　the　imine　bonds.　The　MTT　assays　implied　that　the　hydrogels　were　non-cytotoxic　on　mice　bone　marrow　mesenchymal　stem　cells.　Therefore,　the　hydrogels　showed　potential　application　in　biomedical　fields,　and　consequently　further　work　was　performed　using　the　self-healing　hydrogels　as　drug　carriers　in　in　vitro/vivo　antitumor　studies.　©　2017　The　Royal　Society　of　Chemistry.