Abstract:Objective To isolate macrophages from rhizomys pruinosus, and provide methods for preliminary identification of macrophages based on morphological and phagocytic characteristics; To observe the phagocytosis of different sources of macrophages to Penicillium marneffei conidia and detect the phagocytic rate,which will provide initial experimental evidence of the correlation between Penicillium marneffei, Penicilliosis marneffei and rhizomyidae. Methods Different methods were used to isolate macrophages from alveoli, peritoneum and spleen of rhizomys pruinosus. The cell morphology was observed by Wright-Giemsa staining and transmission electron microscopy. By transmission electron microscopy, the phagocytosis of different sources of macrophages to Penicillium marneffei conidia and the changes of the surface and intracellular structure of macrophages before and after phagocytosis were observed. The phagocytic rate was detected by flow cytometry. Results Different sources of primary macrophages of rhizomys pruinosus were isolated. The isolated cells had the characteristic of macrophages in morphology which have been confirmed by light microscope (Wright-Giemsa staining) and electron microscope. At the same time, the number of cell surface projection increased and became thicker. However, the number and structure of mitochondria, endocytoplasmic reticulum and lysosome did not change after the phagocytosis. The structure of Penicillium marneffei conidia had no change, either. The phagocytic rate of alveolar, peritoneal, spleen macrophages to Penicillium marneffei conidia were 75.09%±6.92%、83.36%±0.87%、78.84%±3.73%, respectively, with statistically significant difference among the three groups (P<0.05). Conclusions The macrophages isolated from alveoli, peritoneum and spleen of rhizomys pruinosus have good phagocytosis to Penicillium marneffei conidia. No lethal effect of the rhizomys pruinosus macrophages on Penicillium marneffei conidia was observed by transmission electron microscopy.