关键词: 胰腺干细胞, 单克隆, 形态, 表达特征, 免疫组织化学, 反转录-聚合酶链式反应, 人

Abstract: Objective Cell morphology and expression features of the monoclonal human pancreatic stem cell line were studied. Methods Pancreatic tissues were taken from ten abortive fetuses of 4-5 month-old by sterile procedures, and digested with collagenase. Primary pancreatic cells were cultured in Dulbecco’s Modified Eagles’s Medium. These cells were further digested with trypsin for propagation. Pancreatic stem cells were gradually purified during generations. The monoclonl human pancreatic stem cells were selected by clone-ring, and further proliferated. Cells which had better viability were preserved in liquid nitrogen. Morphological features of the monoclonal human pancreatic stem cells were observed by hematoxylin-eosin and giemsa staining. Ultrastructure was analyzed by transmission electron microscope. Expression characteristics were demonstrated by immunocytochemistry staining and reverse transcription-polymerase chain reaction (RT-PCR). Results A monoclonal human pancreatic stem cell line which was derived from a male abortive fetus of 4 month-old was established, and had been passed though 50 generations. More than 1×10SUP>9/SUP> monoclonal human pancreatic stem cells were cryo-preserved in liquid nitrogen. The monoclonal human pancreatic stem cell had a polygonal epithelioid morphology with a round or oval caryon and mono or double or poly-nucleolus. The ratio of caryon and cytoplasm was large. There were underdeveloped endoplasmic reticulum, mitochondria, and no excretory granule in cytoplasm. These cells were positive for the pancreatic and duodenal homeobox factor 1 (pdx1), glucagon, nestin and cytokeratin 19 (CK19), and negative for the insulin, cyclin-dependent 34 (CD34), CD44 and CD45 protein expression. Also, transcription factors of the pdx1, glucagon and nestin were expressed. Conclusion A monoclonal human pancreatic stem cell line which co-expressed the pdx1, glucagon, nestin and CK19 proteins was established and identified for the fir

Key words: Pancreatic stem cell, Monoclone, Morphology, Expression, Immunohistochemistry, RT-PCR, Human