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The Dissociation and Culture of Bovine Fibroblasts in Vitro
Ye Rong, Xue-Jin Chen1, Li-Guo Yang2*
College of Life Science, Soochow University, Suzhou 215007, China; 1The Research Center for Development Biology, Shanghai Second Medical University, Shanghai 200025, China; 2College of Animal Science and Technology, Huazhong Agricult
Abstract: The experiment were studied on the dissociation， culture and purification of bovine fetus fibroblasts and skin fibroblasts. The attachment and monolayer cells could be obtained by primary culture with bovine fetus and skin tissue. More fibroblast cells were obtained by 2.5 g/L trypsin+1 mmol/L EDTA and 5 g/L collagenase I digestion bovine skin tisssue than by 2.5 g/L trypsin+1 mmol/L EDTA (P<0.01)， but digestion fetus tissue by 2.5 g/L trypsin+1 mmol/L EDTA got a higher yield of cells than that of skin tissue (P<0.01). The growth curves of fetus and skin fibroblast are normal and similarity， although the death rate of cells increased with the fibroblast yield increase. The death rate of attachment cells digested by 2.5 g/L trypsin+1 mmol/L EDTA was significantly higher than that by 0.5 g/L trypsin+0.53 mmol/L EDTA (P<0.05)， and the attachment rate of the former was significantly lower than the latter (P<0.05) with 24 h culture. Homogeneous fibroblasts could be obtained by 0.5 g/L trypsin digestion of fibroblasts with small part of epithelial cells in it and by passed 2-3 subcultures.
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