Morphological Study And Regression Analysis About In Vitro Phagocytosis Of Apoptotic Granulosa Cells By Monocytes In Yak

The present study firstly established a method for quantitative measurement of phagocytosis in mouse. Then the further research has been carried out about the morphologic change of phagocytosis in yak.On the mouse, the phagocytic process of apoptotic granulosa cells by monocytes was imitated in vitro. Monocytes and granulosa cells were isolated from Kunming mice and cultured. Granulosa cells were induced to apoptosis by garlic, and then co-cultured with monocytes. At different time points (1 h, 2 h, 3 h, 4 h, 5 h), co-cultured cells were assessed by Wright's staining. The results showed that at the beginning of morphological changes in apoptotic granulosa cells, monocytes captured the apoptotic cells. Meanwhile, the apoptosis of granulosa cells were continuing. Debris was found in phagocytic vacuole. At the point of 3 h after co-culture, the ratio between the number of monocytes which attached apoptotic granulosa cells and that which engulfed the apoptotic cells was close to one. Namely, half of monocytes were in the state of recognition and half were in a state of engulfment. The author suggested to name it as 'half phagocytic period'. Regression analysis showed that the equation of linear regression was y = -0.247x + 1.644, R~2=0.912, F=31.095, P=0.011 (<0.05), T= -5.576, P=0.011 (<0.05).On the yak, according to the mode of mouse above, the phagocytic process of apoptotic granulosa cells by monocytes was also imitated in vitro. At different time points (1 h, 2 h, 3 h, 4 h, 5 h), co-cultured cells were assessed by Wright's staining. In addition, the process was observed through scanning electron microscope (SEM) and transmission electron microscope (TEM). The results showed that the morphological changes under microscope were same with that of mouse in the main. At the point of 4 h after co-culture, the ratio between the number of monocytes which attached apoptotic granulosa cells and that which engulfed the apoptotic cells was close to one. Regression analysis showed that the equation of linear regression was y = -0.540x + 2.958, R~2 = 0.937, F=44.766, P=0.007 (<0.01), T= -6.691, P=0.007 (<0.01). Under the SEM and TEM, the apoptosis and phagocytosis were apparent.In conclusion, the present mode of phagocytosis can be used as a method of quantitative assay, which could analyze some effective factors such as medicines that could enhance or restrain phagocytosis. The mode of mouse fit on the study of yak. The result of half phagocytic period and morphological changes enlarged the understanding of apoptosis and phagocytosis, providing base for further research. Based on the mode, the ability of phagocytosis could be studied on other animals. In addition, it was notable that effect of fluorochrome stain could be got through picture processing of 'Invert' on the pictures.