[Quantitative measurement of in vitro phagocytosis of apoptotic granulosa cells by monocytes in mice.].
To establish a method for quantitative measurement of phagocytosis, 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 observed by microscope after 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 progressing. Debris was found in phagocytic vacuole. At the point of 3 h after co-culture, the ratio of monocytes which attached to apoptotic granulosa cells to those which engulfed the apoptotic cells was close to one. Namely, half of monocytes were in the state of recognition and half were in the state of engulfment, and this time point was named as 'half phagocytic period'. Regression analysis showed that the equation of linear regression was y = -0.247x +1.644 (y represents Attachment/Engulfment ratio, x represents co-culture time), R(2)=0.912, F=31.095, P=0.011 (<0.05), T= -5.576, P=0.011 (<0.05). In conclusion, the present mode of phagocytosis in vitro can be used as a method to quantitatively assay some effective factors such as medicines which could enhance or restrain phagocytosis.
Zhao
Hua-Shan
HS
College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, China.
Yu
Si-Jiu
SJ
Zhao
Qiang
Q
D000818:Animals; D017209:Apoptosis; D018920:Coculture Techniques; D005260:Female; D006107:Granulosa Cells / Q000166:cytology*; D051379:Mice; D009000:Monocytes / Q000166:cytology*; D010587:Phagocytosis