Surface markers for the murine oval cell response

Abstract

The biology of progenitor activation in the liver is of considerable medical and scientific interest. The powerful genetic tools available for the mouse make it an ideal model system to study this complex process involving many different cell types. However, reagents for the isolation and study of distinct hepatic subpopulations have been quite limited compared to those available for hematopoietic cells. To produce cell surface reactive reagents more specific for the oval cell response, we generated

Hippo signaling is a potent in vivo growth and tumor suppressor pathway in the mammalian liver

“…The result was statistically analyzed by Student's t test. Immunofluorescent staining was performed as previously described (21). Briefly, freshly isolated liver tissues were embedded in optimal cutting temperature (OCT) embedding medium without fixation and samples were frozen in an isopentane/liquid nitrogen slurry.…”

“…At present no single antibody can unambiguously define murine oval cells, likely due to their heterogeneity. However, most murine oval cells are positive for ductal markers such as M1C1-1C3 and many stain with the OC2-1D11 and A6 antibodies (21,22). Using this panel of antibodies, we consistently observe intense staining of periductal cells in mst1/2 mutant livers, indicating that mst1/2 signaling is required to repress oval cell induction and proliferation.…”

Flow Cytometric Sorting of Cells from Solid Tissue – Reagent Development and Application

“…trypsin) to disperse tightlyassociated cells. Specific protocols successfully used for tissues such as mouse liver (Dorrell et al, 2008b), and human pancreas (Dorrell et al, 2008a) are described in the next section.…”

“…Following all of these treatments little or no undigested material should remain. At each stage, it is important to collect dissociated cells by passing them through a 40μm strainer and to store them on ice without further exposure to enzyme (Dorrell et al, 2008b;Klaunig et al, 1981).…”

“…Under these conditions, cells were sorted at between 2,000 and 4,000 events per second with over 98% purity (Dorrell et al, 2008b). When sorting mouse hepatocytes, which are large and fragile, the optimal nozzle for highest post-sort viability is the 150 μm nozzle at 4.5 PSI (Duncan et al, 2010).…”