Patent ID: 7824912
Filing Date: 2010-11-02
Classification: C07K,C12N,G01N

Abstract:
1. A method for deriving clonal cell lines of somatic liver stem cells comprising: a) plating at confluent cell density a starting liver cell population containing a population of somatic liver stem cells; b) culturing the starting liver cell population containing a population of somatic liver stem cells at confluent cell density of step a) in a culture media with reduced serum; c) adding to the culture media of step b) growth factors TGF-β and EGF, and a guanine nucleotide precursor having a xanthine nucleus, wherein said guanine nucleotide precursor having a xanthine nucleus is present in an amount of 50 μM-5 mM; d) culturing the starting liver cell population of step c) for a time sufficient to permit TGF-β and EGF to induce differentiation and growth arrest of non-stem cells and permit stem cell growth; e) expanding the cells of step d) in the culture media comprising 50 μM-5 mM of a guanine nucleotide precursor having a xanthine nucleus; f) growing individual cell colonies from the cells of step e) in the culture media comprising 50 μM-5 mM of a guanine nucleotide precursor having a xanthine nucleus, and transferring said individual colonies to single wells of a culture plate; and g) culturing and expanding said individual cell colonies that were isolated in step f) in the culture media comprising 50 μM-5 mM of a guanine nucleotide precursor having a xanthine nucleus resulting in expansion of said isolated cells thereby deriving clonal cell lines of somatic liver stem cells.