昆明小鼠胰腺导管上皮样细胞向胰岛样细胞的诱导分化

目的研究正常昆明小鼠胰腺导管上皮样细胞向胰岛样细胞分化的诱导条件。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、48、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。在此条件下培养并传代。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、5、7、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样细胞在高糖刺激下分泌胰岛素的功能。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。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样细胞方向分化。并且该胰岛样细胞在高糖刺激下具有分泌胰岛素的功能。

The Induced Differentiation from Pancreas Ductal Epitheliod Cells to Islet-like Cells of KM Mice

Objective Study the induced condition of the differentiation from the pancreas ductal epitheliod cells to islet-like cells of the normal KM mice. Method Take the pancreas ductal epitheliod cells of normal KM mice for primary culture. Remove the glandular cell by change the medium consecutively at 24 h,48 h,72 h cause of the different adherence time. Remove the fibroblast by use of the medium with 2% FBS and make the pancreas ductal epithelioid cell which can grow in medium free of serum gain dominance. Passage the cell at this condition and induce the third generation differentiate by use of the medium without serum. Take the cell induced for 3 d,5 d,7 d,14 d to identification by immunocytochemistry stain. Detect the ability of insulin-secretion of induced isletlike cells with high glucose condition by ELISA. Result The pancreas ductal epithelioid cells induced with medium free of serum for 7 d start differentiating to islet-like cells that positive stain with anti-insulin antibody. The positive cell mult obviously at 14 d following inducing. And these cells can secret insulin in condition of high glucose. Conclusion The pancreas ductal epithelioid cell can differentiate to islet-like cell by induced with medium free of serum.