摘要
目的 完成胰腺导管上皮细胞的原代及传代培养 ,建立能够体外长期培养的胰腺导管上皮细胞系。 方法 用含有胶原酶 IA型的消化液消化分离胎儿胰腺组织为细小、均匀的细胞团进行接种 ,接种的细胞团用含10 %胎牛血清、4.0 m mol/L谷氨酰胺、0 .0 1%大豆胰酶抑制剂、0 .0 2 %牛血清白蛋白、5 .0 m g/L牛脑垂体提取物、2 .5× 10 - 3m g/L表皮生长因子、2 5 .0× 10 - 2 m g/L霍乱毒素、5 .0 mg/L胰岛素、10 .0 mg/L转铁蛋白、1.0 m g/L地塞米松、5 0 m g/L庆大霉素的完全培养基培养 2 4h后 ,换含 2 %胎牛血清的完全培养基继续培养 ,在细胞团达到 80 %～ 90 %的细胞汇合时 ,1∶ 2传代培养。 结果 获得的原代培养细胞不含有淀粉酶 ,表达细胞角蛋白 ,可初步认定为胰腺导管上皮细胞。原代培养的胰腺导管上皮细胞已传代培养到第 3代。 结论 我们的原代培养和传代培养的方法、条件适宜于胰腺导管上皮体外细胞培养。
Objective\ To establish a normal pancreatic ductal epithelial cell line for further study. Methods The human fetal pancreas was dispersed with the solution containing collagenase type IA. Then cells were maintained in a complete media consisting of 10% fetal bovine serum(FBS), 4 0mmol/L glutamine, 0 01% soybean trypsin inhibitor, 0 02% bovine serum albumin, 5 0mg/L bovine pituitary extract, 2 5×10 -3 mg/L epithelial growth factor, 25 0×10 -2 mg/L cholera toxin, 5 0mg/L insulin, 10 0mg/L transferrin, 1 0mg/L dexamethasone, and 50mg/L gentamycin. Twenty\|four hours later, they were cultured successively in a complete media of 2% FBS. When the cultured cells reached 80%\|90% confluence, they were subcultured in 1∶2. Results\ Expression of cytokeratin and absence in amylase proved that the cultures were pancreatic ductal cells. These pancreatic ductal cells had been subcultured for three generations.Conclusion\ Our culture methods and conditions used in the experiments are suitable for the primaryculture and subculture of human pancreatic ductal cells.
出处
《解剖学报》
CAS
CSCD
北大核心
2000年第2期180-182,I010,共4页
Acta Anatomica Sinica
