犬胆囊上皮细胞的分离及培养

Separation and culture of gallbladder epithelial cells in dogs

目的 探讨胆囊上皮细胞的分离方法和生长条件 ,为研究胆囊的生理功能和相关疾病奠定基础。方法 采有Ⅳ型胶原酶分离细胞 ,Ⅳ型胶原和小牛血清包被培养皿 ,DMEM/HamF12培养基联合培养 ,观察细胞形态和生长曲线 ,并用胆囊上皮细胞特异性标记物角蛋白CK19鉴定细胞。结果 每个犬胆囊可分离得到 5 10× 10 6个胆囊上皮细胞 ,接种一周内生长旺盛 ,第 3 ,4d达到峰值 ,呈现扁平多角形成片贴壁状态。结论 Ⅳ型胶原酶消化法能成功分离家兔胆囊上皮细胞 ,Ⅳ型胶原 /小牛血清可促进细胞贴附 ,DMEM /HamF12培养基可提供较为满意的生长条件。

Objective To investigate the separation methods and growth conditions of gallbladder epithelial cells,and establish the ground -work for the research of physiologic function of gallbladder and the related diseases.Methods The epithelial cells were separated by collagenase typeⅣ and cultured under the combinated substrate of DMEM/HamF12.The culture dishes were coated with collagen type Ⅳ and bovine calf serum initially.Then the morphologic changes of the cells was observed and the growth curve of the cells was made. Results A total of 5-10×10 6 cells were obtained from each dog gallbladder.These cells grew vigorously within one week and reached the maximum at the 3rd and 4th days.And these cells presented flat,and multangular,and attached flakily with the dishes.Conclusion The method of collagenase type Ⅳ digestion can successfully separate the dog gallbladder epithelial cells.Collagen type Ⅳ and bovine calf serum can promote the attachment of the cells.DMEM/HamF12 can provide satisfactory growth condition for gallbladder epithelial cells.