胎猪胰岛源胰腺干细胞分离培养与诱导分化试验.pdfVIP

中国农业科学 2007,40(3):582-587 Scientia Agricultura Sinica 胎猪胰岛源胰腺干细胞分离培养与诱导分化试验 冯若鹏，张慧茹，窦忠英 （西北农林科技大学，国家干细胞工程技术研究中心陕西省分中心, 杨凌 712100 ） 摘要：【目的】分离培养胎猪胰腺干细胞对其进行功能检测。【方法】用先悬浮后贴壁的方法从胎猪胰岛获得 胰腺干细胞，用免疫组化方法鉴定该细胞，以 MTT 法测定其不同代次的生长活性，诱导该细胞成为胰腺内分泌细 胞并检测其功能。【结果】分离得到的细胞不表达nestin，而表达导管上皮细胞标志ck-19、平滑肌标志α-actin, 其在体外的生长行为有类似胚胎干细胞样生长形态，细胞体外增殖活力很强，现已传至18代，细胞经诱导后分化 为分泌胰岛素的胰腺内分泌细胞，并且伴随着细胞超微结构的改变。【结论】通过本方法可以分离到胎猪胰腺干/ 祖细胞，经诱导分化可形成有功能的胰岛样细胞团并释放胰岛素。 关键词：胎猪；胰岛；胰腺干细胞；培养；诱导分化 Iolation, Culture, and Induced Differentiation of Fetal Porcine IPSCS FENG Ruo-peng ，ZHANG Hui-ru ，DOU Zhong-ying （Shaanxi Branch Center of China Stem Cell Engineering and Technology Research Center, Northwest Sci-Tech University of Agriculture and Forestry, Yangling 712100 ） Abstract ：【Objective 】The object was to isolate and culture the porcine pancreatic stem cells and investigate its function. 【Method 】Scientists isolated the fetal porcine pancreatic stem cells by suspending-adhering. The isolated cells were identified by immunohisto- chemical staining. Scientists then measured the growth vigor of these cells through MTT method in vitro and induced these cells to differentiate into endocrine cells and detect its function. 【Results 】The isolated IPSCS did not express nestin but expressed the marker of ductal cells and smooth muscle-CK-19 and of α-actin. The growth characteristic of these cells resembled that of the ES cells.These cells which were subcultured 18 times possessed a strong proliferative ability. The cells could excrete insulin and accompany with the ultrastructure alteration after induction. 【Conclusion 】Porcine pancreatic stem cells can be isolated by this method, these cells can be induced to form islet-like clusters and s