# US 12,110,507 B2 · Method for preparing functional hepatic progenitor cells or hepatic cells, or functional small intestinal epithelial progenitor cells or small intestinal epithelial cells > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,110,507 B2 - Document ID: US12110507B2 - Status: Granted - Kind: B2 - Application number: 16/958,118 - Filed: 2018-12-27 - Published: 2020-10-29 - Granted: 2024-10-08 - Earliest claimed: 2017-12-27 - Assignees: Akihiro Umezawa - Applicants: Akihiro Umezawa - Inventors: Akihiro Umezawa; Tohru Kiyono - Web page: https://patentgrep.com/patent/US12110507B2 ## CPC - C12N 5/0672: {Stem cells; Progenitor cells; Precursor cells; Oval cells} - A61K 35/407: Liver; Hepatocytes - A61L 27/38: {containing added animal cells (organs or tissue containing native cells A61L27/36)} - A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics - C12N 5/068: {Stem cells; Progenitors} - C12N 15/00: Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor (mutants or genetically engineered microorganisms, per se C12N1/00, C12N5/00, C12N7/00; new plants per se A01H; plant reproduction by tissue culture techniques A01H4/00; new animals per se A01K67/00; use of medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases, gene therapy A61K48/00) - G01N 33/5067: {Liver cells} - C12N 2500/05: Inorganic components - C12N 2500/38: Vitamins - C12N 2501/105: Insulin-like growth factors [IGF] - C12N 2501/115: Basic fibroblast growth factor (bFGF, FGF-2) - C12N 2506/02: from embryonic cells - C12N 2506/45: from artificially induced pluripotent stem cells - C12N 2510/00: Genetically modified cells - C12N 2510/04: Immortalised cells - C12N 2533/54: Collagen; Gelatin - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61L: METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES (preservation of bodies or disinfecting characterised by the agents employed A01N; preserving, e.g. sterilising, food or foodstuffs A23; preparations for medical, dental or toiletry purposes A61K) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract Provided is a method for preparing functional hepatic (progenitor) cells or functional small intestinal epithelial (progenitor) cells, comprising the step of culturing an isolated cell population comprising hepatic (progenitor) cells or small intestinal epithelial (progenitor) cells in the presence of an antibiotic. Also, provided is a substantially homogeneous isolated cell population comprising functional hepatic progenitor cells, wherein an expression level of CYP3A4 in the functional hepatic progenitor cells is increased by at least 5 times the expression level thereof in a HepaRG® cell line. ## Claims ### Claim 1 A method for preparing functional hepatic progenitor cells or functional hepatic cells, or functional small intestinal epithelial progenitor cells or functional small intestinal epithelial cells, the method comprising the step of culturing an isolated cell population comprising hepatic progenitor cells or hepatic cells, or small intestinal epithelial progenitor cells or small intestinal epithelial cells in the presence of 0.1 to 100 μg/ml puromycin and 5 to 15 mg/ml ammonia, wherein the hepatic progenitor cells or hepatic cells, or the small intestinal epithelial progenitor cells or small intestinal epithelial cells do not comprise an exogenous resistance gene for the antibiotic. ### Claim 2 (depends on 1) The method according to claim 1, wherein the isolated cell population is a primary culture derived from a hepatic tissue or a small intestinal epithelial tissue. ### Claim 3 (depends on 1) The method according to claim 1, wherein the isolated cell population is differentiated from stem cells. ### Claim 4 (depends on 3) The method according to claim 3, wherein the stem cells are embryonic stem (ES) cells. ### Claim 5 (depends on 3) The method according to claim 3, wherein the stem cells are induced pluripotent stem (iPS) cells. ### Claim 6 (depends on 5) The method according to claim 5, wherein the iPS cells are derived from a healthy individual. ### Claim 7 (depends on 5) The method according to claim 5, wherein the iPS cells are derived from a drug-induced liver injury patient or a drug-induced small intestinal injury patient. ### Claim 8 (depends on 1) The method according to claim 1, wherein the culture step is performed in the presence of a hepatic differentiation inducing factor. ### Claim 9 (depends on 8) The method according to claim 8, wherein the hepatic differentiation inducing factor is selected from the group consisting of a cytokine, a cell growth factor, a ROCK inhibitor, a MAPK inhibitor, an ALK inhibitor and an extracellular matrix. ### Claim 10 (depends on 1) A stable functional hepatic progenitor cell line or a stable functional hepatic cell line obtained by a method according to claim 1, wherein an expression level of CYP3A4 in the functional hepatic progenitor cells is increased by at least 5 times the expression level thereof in a HepaRG cell line, an immortalized mature hepatic cell line, or a hepatic cell line obtained by inducing differentiation without selection based on puromycin. ### Claim 11 (depends on 1) A stable functional small intestinal epithelial progenitor cell line or a stable functional small intestinal epithelial cell line obtained by a method according to claim 1, wherein an expression level of CYP3A4 in the functional hepatic progenitor cells is increased by at least 5 times the expression level thereof in a HepaRG cell line, an immortalized mature hepatic cell line, or a hepatic cell line obtained by inducing differentiation without selection based on puromycin.