# US 12,215,353 B2 · Plated hepatocytes and preparation and uses thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,215,353 B2 - Document ID: US12215353B2 - Status: Granted - Kind: B2 - Application number: 18/074,837 - Filed: 2022-12-05 - Published: 2023-03-30 - Granted: 2025-02-04 - Earliest claimed: 2017-09-22 - Assignees: LifeNet Health - Applicants: LifeNet Health - Inventors: Jung Bok Lee; Angela Murchison; Edward LeCluyse; Jingsong Chen - Web page: https://patentgrep.com/patent/US12215353B2 ## CPC - C12N 5/067: {Hepatocytes} - G01N 33/5067: {Liver cells} - C12N 2502/1323: Adult fibroblasts - C12N 2502/28: Vascular endothelial cells - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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 The present invention provides a product comprising plated human hepatocytes on a surface and at least some of the plated hepatocytes are in one or more hepatocyte clusters on feeder cells, which are attached to the surface. A method of preparing plated human hepatocytes is also provided. The preparation method comprises applying human hepatocytes to a surface in the presence of feeder cells, co-culturing the applied hepatocytes with the feeder cells, and forming one or more hepatocyte clusters by the co-cultured hepatocytes on the feeder cells, which are attached to the surface. The plated hepatocytes may be used for various purposes, including the preparation of a hepatitis B virus (HBV) infected hepatocyte culture model and drug testing. ## Claims ### Claim 1 A product comprising plated hepatocytes on a surface, wherein at least 70% of the plated hepatocytes are in one or more hepatocyte clusters on feeder cells, wherein the feeder cells are endothelial cells and fibroblasts and are attached to the surface, wherein the plated hepatocytes are obtained from one human donor, one non-human donor, multiple human donors, or multiple non-human donors; the endothelial cells are obtained from one human donor, one non-human donor, multiple human donors, or multiple non-human donors; and the fibroblasts are obtained from one human donor, one non-human donor, multiple human donors, or multiple non-human donors; and wherein the plated hepatocytes, the endothelial cells and the fibroblasts have a cell ratio from 3:1:1 to 24:1:1. ### Claim 2 (depends on 1) The product of claim 1, wherein the plated hepatocytes are obtained from one non-human donor, the endothelial cells are obtained from multiple human donors and the fibroblasts are obtained from multiple human donors. ### Claim 3 (depends on 1) The product of claim 1, wherein the plated hepatocytes are obtained from multiple human donors, the endothelial cells are obtained from multiple non-human donors and the fibroblasts are obtained from multiple non-human donors. ### Claim 4 (depends on 1) The product of claim 1, wherein the plated hepatocytes, the endothelial cells and the fibroblasts are obtained from multiple non-human donors. ### Claim 5 (depends on 1) The product of claim 1, wherein the fibroblasts do not proliferate. ### Claim 6 (depends on 1) The product of claim 1, wherein the fibroblasts are not immortal. ### Claim 7 (depends on 1) The product of claim 1, wherein the plated hepatocytes are obtained from one human donor, the endothelial cells are obtained from one human donor, and the fibroblasts are obtained from one human donor. ### Claim 8 (depends on 1) The product of claim 1, wherein the plated hepatocytes are obtained from multiple human donors, the endothelial cells are obtained from multiple human donors and the feeder cells are obtained from multiple human donors. ### Claim 9 A product comprising plated hepatocytes on a surface, wherein at least 70% of the plated hepatocytes are in one or more hepatocyte clusters on feeder cells, wherein the feeder cells are endothelial cells and fibroblasts and are attached to the surface, wherein the plated hepatocytes are obtained from one human donor, the endothelial cells are obtained from multiple human donors, and the fibroblasts are obtained from multiple human donors. ### Claim 10 (depends on 1) The product of claim 1, wherein the plated hepatocytes are obtained from one human donor, the endothelial cells are obtained from one non-human donor, and the fibroblasts are obtained from one non-human donor. ### Claim 11 (depends on 1) The product of claim 1, wherein the plated hepatocytes are obtained from one human donor, the endothelial cells are obtained from multiple non-human donors, and the fibroblasts are obtained from multiple non-human donors. ### Claim 12 (depends on 1) The product of claim 1, wherein the plated hepatocytes are obtained from multiple human donors, the endothelial cells are obtained from one human donor, and the fibroblasts are obtained from one human donor. ### Claim 13 (depends on 1) The product of claim 1, wherein the plated hepatocytes are obtained from multiple human donors, the endothelial cells are obtained from one non-human donor, and the endothelial cells are obtained from one non-human donor. ### Claim 14 (depends on 1) The product of claim 1, wherein the plated hepatocytes are obtained from one non-human donor, the endothelial cells are obtained from one human donor, and the fibroblasts are obtained from one human donor. ### Claim 15 (depends on 1) The product of claim 1, wherein the plated hepatocytes are obtained from one non-human donor, the endothelial cells are obtained from one non-human donor, and the fibroblasts are obtained from one non-human donor. ### Claim 16 (depends on 1) The product of claim 1, wherein the plated hepatocytes are obtained from one non-human donor, the endothelial cells are obtained from multiple non-human donors, and the fibroblasts are obtained from multiple non-human donors. ### Claim 17 (depends on 1) The product of claim 1, wherein the plated hepatocytes are obtained from multiple non-human donors, the endothelial cells are obtained from one human donor, and the fibroblasts are obtained from one human donor. ### Claim 18 (depends on 1) The product of claim 1, wherein the plated hepatocytes are obtained from multiple non-human donors, the endothelial cells are obtained from multiple human donors, and the fibroblasts are obtained from multiple human donors. ### Claim 19 (depends on 1) The product of claim 1, wherein the plated hepatocytes are obtained from multiple non-human donors, the endothelial cells are obtained from one non-human donor, and the fibroblasts are obtained from one non-human donor.