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10.1038/nprot.2015.017 http://scihub22266oqcxt.onion/10.1038/nprot.2015.017 C4681437!4681437 !25612230     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=25612230 &cmd=llinks): Failed to open stream: HTTP request failed! 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High-yield and high-purity isolation of hepatic stellate cells from normal and fibrotic mouse livers |pdf=|usr=}}{{25612230 }} gab.com Text High-yield and high-purity isolation of hepatic stellate cells from normal and fibrotic mouse livers JO: Nat Protoc http://www.kidney.de/pget.php?&tw=1&pmid=25612230 #FOAvip Hepatic stellate cells (HSCs) have been identified as the main fibrogenic cell type in the liver. Hence, efforts to understand hepatic fibrogenesis and to develop treatment strategies have focused on this cell type. HSC isolation, originally developed in rats, has subsequently been adapted to mice, thus allowing the study of fibrogenesis by genetic approaches in transgenic mice. However, mouse HSC isolation is commonly hampered by low yield and purity. Here we present an easy-to-perform protocol for high-purity and high-yield isolation of quiescent and activated HSCs in mice, based on retrograde pronase-collagenase perfusion of the liver and subsequent density-gradient centrifugation. We describe an optional add-on protocol for ultrapure HSC isolation from normal and fibrotic livers via subsequent flow cytometric sorting, thus providing a validated method to determine gene expression changes during HSC activation devoid of cell culture artifacts or contamination with other cells. The described isolation procedure takes ?4 h to complete. Twit Text FOAVip High-yield and high-purity isolation of hepatic stellate cells from normal and fibrotic mouse livers ????Nat Protoc http://www.kidney.de/pget.php?&tw=1&pmid=25612230 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=25612230 #FOAvip English Wikipedia <ref>{{Cite pmid|25612230 }}</ref> High-yield and high-purity isolation of hepatic stellate cells from normal and fibrotic mouse livers #MMPMID25612230 Mederacke I ; Dapito DH ; Affò S ; Uchinami H ; Schwabe RF Nat Protoc 2015[Feb]; 10 (2 ): 305-15 PMID25612230 show ga Hepatic stellate cells (HSCs) have been identified as the main fibrogenic cell type in the liver. Hence, efforts to understand hepatic fibrogenesis and to develop treatment strategies have focused on this cell type. HSC isolation, originally developed in rats, has subsequently been adapted to mice, thus allowing the study of fibrogenesis by genetic approaches in transgenic mice. However, mouse HSC isolation is commonly hampered by low yield and purity. Here we present an easy-to-perform protocol for high-purity and high-yield isolation of quiescent and activated HSCs in mice, based on retrograde pronase-collagenase perfusion of the liver and subsequent density-gradient centrifugation. We describe an optional add-on protocol for ultrapure HSC isolation from normal and fibrotic livers via subsequent flow cytometric sorting, thus providing a validated method to determine gene expression changes during HSC activation devoid of cell culture artifacts or contamination with other cells. The described isolation procedure takes ?4 h to complete. |*Hepatic Stellate Cells [MESH] |Animals [MESH] |Centrifugation, Density Gradient [MESH] |Collagenases/chemistry [MESH] |Cytological Techniques/instrumentation/*methods [MESH] |Flow Cytometry [MESH] |Liver Cirrhosis/*pathology [MESH] |Liver/cytology [MESH] |Mice, Transgenic [MESH]High-yield and high-purity isolation of hepatic stellate cells from no(epmc).pdf DeepDyve Pubget Overpricing