Objective:　To　establish　a　simple　method　for　isolation,　purification　and　cultivation　of　primary　retinal　microvascular　pericytes　(RMPs)　from　mice.Methods:　Retinas　were　isolated　from　mice　following　with　mechanical　morcel,　enzymatic　digestion　and　filtration.The　retinal　fragments　were　incubated　with　low　glucose　DMEM　with　20%　fetal　bovine　serum　after　24　hours　pre-incubation.Differential　digestion　was　used　for　purification　of　primary　RMPs.Morphological　examination　of　cells　was　performed　by　phase　contrast　microscopy,　and　further　characterization　was　analyzed　by　immunocytochemistry.Functional　assay　was　evaluated　by　the　pericytes-endothelial　cells　(ECs)　co-culture　system.The　treatment　and　use　of　experimental　animals　followed　the　regulations　on　the　administration　of　experimental　animals　promulgated　by　the　state　science　and　technology　commission.Results:　Cells　migrated　out　of　fragments　after　24　hours　of　incubation,　and　developed　into　small　or　large　colonies　gradually.The　cells　and　their　subpassages　presented　typical　pericyte　morphology　with　large　irregular　triangular　cell　bodies　and　multiple　long　processes.No　contact　inhibition　was　observed.Most　cells　uniformly　expressed　the　cellular　markers　α-smooth　muscle　actin　(α-SMA)　and　platelet-derived　growth　factor　receptor-β　(PDGFR-β),　a　few　cells　expressed　the　cellular　markers　glial　fibrillary　acidic　protein　(GFAP),　but　no　cell　expressed　von　Willebrand　factor　(vWF).　The　purity　rate　of　RMPs　was　up　to　97%.In　the　co-culture　system,　RMPs　directly　contacted　with　ECs　to　form　the　capillary-like　cords　in　vitro.Conclusions:　A　simple　method　for　the　isolation,　purification　cultivation　of　mouse　RMPs　is　established,　and　active　RMPs　can　be　readily　obtained　by　this　method.　Copyright　©　2019　by　the　Chinese　Medical　Association.