关键词: 遗传霉素(G418), 成纤维细胞, 细胞空泡化, 许旺细胞, 光学显微镜

Abstract:

BACKGROUND: Geneticin (G418) combined with differential adhesion method and differential detachment method are used to eliminate the contaminated fibroblasts in the purification of Schwann cells. Most of the researches prefer to describe purity of Schwann cells, rather than give a description of cell morphology change and its probable cause during the purification process. 
OBJECTIVE: To describe the pathological changes of fibroblasts through the morphological photos of purified fibroblasts and Schwann cells at different time points taken by optical microscope.
METHODS: Schwann cells were isolated from sciatic nerves of rats by geneticin (G418) combined with differential adhesion method and differential detachment method in primary cultivation. The cultivated cells were divided into two groups. In the experimental group, the cells were transferred to another 6-well plate after once differential adhesion, and the waste product accumulated medium was changed to fresh basal medium every 2 days, the differential detachment method was used to passage. In the control group, cells were given differential detachment once again and transferred to another 6-well tissue culture plate, and the waste product accumulated medium was changed to fresh purification medium every 2 days, the differential detachment method was used to passage. Cytopathologic changes of fibroblasts in different periods were observed by optical microscope.
RESULTS AND CONCLUSION: Cytopathologic changes of fibroblasts could not be observed under optical microscope in early stage of the purification, the proliferation of the fibroblasts was decreased and the cytoplasm vacuolization of fibroblasts was observed obviously after cells were sub-cultured for three times or more (three to four weeks). Fibroblast proliferation was restricted in early stage of the Schwann cells purification process by geneticin (G418), but cytoplasm vacuolization of fibroblasts delayed to the third generation or more.