Liver reserve cells
First Claim
1. Isolated liver reserve cells which retain and are stained by Eosin Y dye when stained successively by a solution comprising Eosin Y dye, and then by a solution comprising both methylene blue and Azure A dyes.
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Abstract
The present invention relates to liver reserve or progenitor cells. In particular, it relates to the isolation, characterization, culturing, and uses of liver reserve cells. Liver reserve cells isolated by density gradient centrifugation can be distinguished from other liver parenchymal cells by their morphology, staining characteristics, high proliferative activity and ability to differentiate in vitro. In long-term cultures described herein, these cells expand in numbers and differentiate into morphologically mature liver parenchymal cells, capable of mediating liver-specific functions. Therefore, isolated liver reserve cells may have a wide range of applications, including, but not limited to, their uses as vehicles of exogenous genes in gene therapy, and/or to replace and reconstitute a destroyed, infected, or genetically deficient mammalian liver by transplantation.
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11 Claims
- 1. Isolated liver reserve cells which retain and are stained by Eosin Y dye when stained successively by a solution comprising Eosin Y dye, and then by a solution comprising both methylene blue and Azure A dyes.
- 7. A cellular composition wherein about 90% of the cells are liver reserve cells which retain and are stained by Eosin Y dye when stained successively by a solution comprising Eosin Y dye, and then by a solution comprising both methylene blue and Azure A dyes.
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10. A method of isolating liver reserve cells comprising subjecting a mixed population of liver cells to 70% Percoll density gradient centrifugation to obtain thereby four zones, the lower two of which contain the liver reserve cells;
- and pooling and centrifuging the cells in the two lower zones against a 25%/50% Percoll discontinuous gradient to obtain thereby an interface zone containing cells having a bouyant density of 1.0381 g/ml.
- View Dependent Claims (11)
Specification