Supplemental Methods GUT Revision 2
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چکیده
LSEC Isolation Liver sinusoidal endothelial cells were isolated from wild type C567BL/6 mice by collagenase perfusion, iodixanol density gradient centrifugation and centrifugal elutriation, as described. Briefly, yields of LSEC are on average 80 million sinusoidal endothelial cells per 10 g liver with viability of more than 95% and purity greater than 96%, as determined by scanning electron microscopy (SEM), fluorescence-activated cell sorter (FACS) for CD31 and CD105, and positive staining for fluorescent acetylated low-density lipoprotein (Dil-Ac-LDL; Biomedical Technologies, Stoughton, MA) and FITC–FSA. FACS on F4/80 and desmin were used to exclude contamination by macrophage/Kupffer cells and HSC. Cells were cultured on rat-tail collagencoated plates (400,000 cells/cm) in Dulbecco's minimal essential medium (DMEM) low glucose with 10% fetal bovine serum (FBS).
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تاریخ انتشار 2014