仔猪胰腺干／祖细胞的生物学特性

分离1日龄长白仔猪的胰腺细胞后，通过显微镜和电镜观察细胞生长形态和超微结构，运用细胞计数法制作细胞生长曲线测定其群体倍增时间，采用免疫组化法测定细胞生长不同时期的表面标志蛋白的表达，用肝细胞因子和尼克酰胺联合诱导其向功能性G细胞分化，并测定了分化细胞对葡萄糖的反应能力。结果显示获得的仔猪胰腺祖／干细胞具有多种形态，以神经样细胞克隆生长占优势，电镜超微结构证实其具有核质比高、细胞器不分化的干细胞态特征。试验测得第1代和第3代群体的倍增时间分别为96．0、307．7h，仔猪胰腺祖／干细胞表达胚胎干细胞标志Oct4、SSEA-1、SSEA-4，也表达胰腺干细胞的标志PDX-1、CK7等。仔猪胰腺干／祖细胞的向β细胞诱导分化细胞对22mmol／L葡萄糖的刺激产生强烈的胰岛素分泌反应，胰岛素分泌量可达321．75mIU／L。结果证实仔猪胰腺千／祖细胞具有干细胞和胰腺祖的表面标志和超微结构，同时具有分化成分泌胰岛素β细胞的能力。

Biological characterization of pancreatic stem/ progenitor in neonatal piglets

The object was to evaluate the characteristics of the neonatal piglet pancreatic stem/progenitor cells for the research basic data. The morphology and ultrastructure of pancreatic cells from the neonatal piglet were observed by microscope and electron microscope. The growth curve and double time was determined. The expression of surface marker protein was analyzed by immunohistochemistry （ABC） method in different phase of the cell growth. The cells were treated with hepatocyte growth factor（HGF） and nicotinamide to differentiate them into β-cells which secreted insulin,and the cells were sensitive to the glucose concentration for the insulin secretion. The results showed that the obtained cells have polymorphism of which neuron-like cells and fibro-blasts were superiority. The ultrastructure by electron microscope determined the property of stem cells which were high nucleo-eytoplasmic ratio and petty organ- elles. The double times of colony subcultured first-times and third-times were 96.0 h and 307.7 h respectively. The neonatal piglets pancreatic stem/progenitor cells had expressed the embryonic stem cells markers which were Oct4, SSEA 1,SSEA-4 ,and expressed the pancreatic stem cells markers, such as PDX-1, CKT. The induced cells secreted 321.75μIU/mL insulin by corresponding to 22 mmol/L glucose concentration,which were toward differentiation en- docrine β-cell. To sum up, the neonatal piglets pancreatic stem/progenitor cells possesse the surface markers and ul- trastructure of stem cells which the capability to differentiate the β-cell to secret insulin.