Hepatocytes with high regenerative capacity for liver repair
View Patent ↗The invention provides purified mammalian hybrid hepatocyte (HybHP) cells, compositions comprising HybHP cells, methods for purifying HybHP cells, methods for in vitro culture of HybHP cells, and methods for using HybHP cells to repopulate and/or treat the liver of a subject in need thereof.
1. A method for purifying a hybrid hepatocyte (HybHP) cell from a normal mammalian liver, said method comprising,
a) preparing a single-cell suspension from said normal mammalian liver,
b) combining said single-cell suspension with
i) at least one first antibody that specifically binds to a plasma membrane first protein marker of liver ductal (DC) cells, and
ii) at least one second antibody that specifically binds to a second protein marker of conventional hepatocyte (cHP) cells, wherein said second protein marker of cHP cells is underexpressed in said HybHP cells compared to cHP cells,
wherein said combining is under conditions for specific binding of said at least one first antibody to said first protein marker, and of said at least one second antibody to said second protein marker, and wherein said specific binding produces a first composition that comprises a first antibody-HybHP cell-second antibody conjugate, and
c) isolating said first antibody-HybHP cell-second antibody conjugate from said single-cell suspension, thereby producing a second composition that comprises a purified HybHP cell.
2. A method for purifying a hybrid hepatocyte (HybHP) cell from a normal mammalian liver, said method comprising,
a) preparing a single-cell suspension from said normal mammalian liver,
b) substantially removing ductal cells from said single-cell suspension to obtain a first population of cells that contains conventional hepatocyte (cHP) cells and HybHP cells,
c) combining said first population of cells with at least one first antibody that specifically binds to a plasma membrane first protein marker of liver ductal (DC) cells, wherein said combining is under conditions for specific binding of said at least one first antibody to said first protein marker, and wherein said specific binding produces a first composition that comprises a first-antibody-HybHP cell conjugate, and
d) isolating said first antibody-HybHP cell conjugate from said first population of cells, thereby producing a second population of cells that comprises a purified HybHP cell.