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Modified methods for isolation of pancreatic stellate cells from human and rodent pancreas

The Journal of Biomedical Research 2016年第6期30卷 510-516页

作者： Liangtao Zhao Baobao Cai Zipeng Lu Lei Tian Song Guo Pengfei Wu Dong Qian Qingcheng Xu Kuirong Jiang Yi Miao Pancreas Institute of Nanjing Medical University Nanjing Jiangsu 211166 China Pancreas Center the First Affiliated Hospital of Nanjing Medical UniversityNanjing Jiangsu 210029 China Lab for Department of General Surgery the First Affiliated Hospital of Nanjing Medical UniversityNanjing Jiangsu 210029 China

Primary cultures of pancreatic stellate cells（PSCs） remain an important basis for in vitro ***,effective methods for isolating abundant PSCs are currently *** report on a novel approach to isolating PSCs from normal rat pancreases and human pancreatic ductal adenocarcinoma（PDAC） *** anaesthesia and laparotomy of the rat,a blunt cannula was inserted into the pancreatic duct through the anti-mesentery side of the duodenum,and the pancreas was slowly infused with an enzyme solution until all lobules were fully *** pancreas was then pre-incubated,finely minced and incubated to procure a cell *** were obtained after the cell suspension was filtered,washed and subject to gradient centrifugation with Nycodenz *** human PDAC tissue was finely minced into 1×1×l mm^3 cubes with sharp *** blocks were placed at the bottom of a culture plate with fresh plasma（EDTA-anti-coagulated plasma from the same patient,mixed with CaCL） sprinkled around the *** culture for 5-10 days under appropriate conditions,activated PSCs were *** intraductal perfusion of an enzyme solution simplified the procedure of isolation of rat PSCs,as compared with the multiple injections technique,and a modified outgrowth method significantly shortened the outgrowth time of the activated *** modification in PSC isolation methods significantly increased the isolation efficiency and shortened the culture period,thus facilitating future PSC-related research.

关键词： pancreatic stellate cells isolation modification

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