Objective: isolation and culture of Tibetan miniature pig rib chondrocytes (PCCs), followed by identifying it. Methods: a 3-day-old Tibetan miniature pig was used in this experiment. Both PCCs and mouse rib chondrocytes (MCCs) were isolated by two-step digestion method. The isolated chondrocytes were identified by toluidine blue staining and immunofluorescence for collagen II and aggrecan. Tibetan miniature pig embryonic fibroblasts (PEFs) and mouse embryonic fibroblasts (MEFs) were employed as a negative control. Results: PCCs and MCCs were successfully isolated and cultured, and demonstrated the different cell morphology, whereas toluidine blue staining and immunofluorescence staining confirmed the chondrocytes characteristics of isolated PCCs. Conclusion: The method for isolating, culturing and identifying PCCs of Tibetan miniature pig was successfully established.