昆明小鼠胰腺导管上皮样细胞向胰岛素分泌细胞的诱导分化

发布时间：2019-01-22 19:08

【摘要】： 目的：通过观察原代昆明小鼠胰腺导管上皮样细胞在动态培养条件下的分化状态以及该分化后的细胞分泌胰岛素的功能，探讨正常昆明小鼠胰腺导管上皮样细胞向胰岛素分泌细胞分化的诱导条件、诱导后的胰岛素分泌细胞分泌胰岛素的功能以及分泌的胰岛素的降糖活性，为1型糖尿病的细胞替代治疗提供实验依据。方法：取正常成年昆明小鼠胰腺导管上皮进行原代培养，利用细胞贴壁时间的差异在培养24 h、48 h、72 h连续换液除去腺体细胞，在含2％胎牛血清的培养条件下除去成纤维细胞，，使可以在无血清培养基中生长的胰腺导管上皮样细胞形成优势生长。在此条件下培养、传代并鉴定。取第三代细胞以无血清诱导培养基促使其分化。取诱导后3 d、5 d、7 d、14 d的细胞进行免疫细胞化学染色鉴定。ELISA检测诱导生成的胰岛素分泌细胞在高糖刺激下分泌胰岛素的功能。用高糖培养基刺激诱导生成的胰岛素分泌细胞后离心取上清液注射到正常昆明小鼠腹部皮下，检测所分泌的胰岛素的降糖活性。结果：经过无血清诱导培养基诱导7 d后，胰腺导管上皮样细胞开始向胰岛素分泌细胞分化，胰岛素抗体染色阳性。14 d胰岛素抗体染色阳性细胞明显增多。该细胞在高糖刺激下可以分泌胰岛素。所分泌的胰岛素可以使正常昆明小鼠的血糖下降。结论：昆明小鼠胰腺导管上皮样细胞经过无血清培养基诱导可以向胰岛素分泌细胞方向分化。并且该胰岛素分泌细胞在高糖刺激下具有分泌胰岛素的功能。所分泌的胰岛素具有降糖活性。
[Abstract]:Aim: to observe the differentiation of primary pancreatic ductal epithelioid cells in Kunming mice under dynamic culture and the function of insulin secretion by the differentiated cells. To investigate the inducing conditions of differentiation of pancreatic ductal epithelioid cells into insulin secreting cells in normal Kunming mice, the function of insulin secretion cells and the hypoglycemic activity of insulin secreted by induced insulin secretion cells. To provide experimental evidence for cell replacement therapy of type 1 diabetes. Methods: the pancreatic ductal epithelium of normal adult Kunming mice was cultured in primary culture. The glandular cells were removed from the pancreatic ductal epithelium at 24 h, 48 h and 72 h by cell adhesion time. The fibroblasts were removed under the condition of 2% fetal bovine serum, and the pancreatic ductal epithelioid cells which could grow in serum-free medium formed dominant growth. In this condition, culture, passage and identification. The third generation cells were induced to differentiate by serum-free medium. The immunocytochemical staining was performed on the cells at 3 days, 5 days, 7 days and 14 days after induction. ELISA was used to detect the function of insulin secreting cells induced by high glucose stimulation. The insulin secreting cells induced by high glucose medium were then centrifuged and injected into the abdominal subcutaneous of normal Kunming mice to detect the hypoglycemic activity of the insulin secreted. Results: after induction on serum-free medium for 7 days, pancreatic ductal epithelioid cells began to differentiate into insulin-secreting cells, and insulin antibody staining positive cells increased significantly at 14 days. The cells secrete insulin under high glucose stimulation. The insulin secreted can reduce the blood sugar of normal Kunming mice. Conclusion: pancreatic ductal epithelioid cells of Kunming mice can differentiate into insulin secreting cells induced by serum-free medium. Furthermore, the insulin secreting cells have the function of secreting insulin under high glucose stimulation. The insulin secreted has hypoglycemic activity.
【学位授予单位】：四川大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R329

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