The United States of America As Represented by the Secretary of

The United States of America As Represented by the Department of Health

Katharine H Cole

Curtis C Harris

John F Lechner

Roger R Reddel

Andrea M Pfeifer

Immortalized cell lines derived from normal adult human liver are described which express phenotypic characteristics of normal adult liver epithelial cells. The invention further provides methods of using the immortalized cells to evaluate the cytotoxicity or carcinogenicity of a compound.

Human liver epithelial cell lines

Immortalized cell lines derived from normal adult human liver are described which express phenotypic characteristics of normal adult liver epithelial cells.

The present invention relates to long term multiplication and permanent establishment of a cell line of human liver epithelial cells(hepatocytes). The human liver epithelial cell line is capable of mitotically proliferating and continuously growing in vitro under suitable environmental conditions in suitable culture media. A method of producing an immortalized human liver epithelial cell line is also disclosed. The invention also relates to serum-free cell medium developed to support long term multiplication and permanent establishment of a cell line of human liver epithelial cells. The medium may contain an effective cell growth promoting amount of calcium ions; an effective cell growth promoting amount of glucose; an effective amount of insulin to aid cells in glucose uptake; an effective cell growth promoting amount of hydrocortisone; an effective amount of epidermal growth factor to bind epidermal growth factor receptors on cells; an effective amount of transferrin to increase DNA synthesis in cells; an effective amount of cholera toxin to increase DNA synthesis in cells; an effective amount of triiodothyronine to increase DNA synthesis in cells; and an effective growth promoting amount of mammalian hormones and mitogenic factors, including lipoprotein, cholesterol, phospholipids and fatty acids.

Human liver epithelial cell line and culture media therefor

The present invention relates to cell medium developed to support long term multiplication and permanent establishment of a cell line of human liver epithelial cells. The medium may contain an effective cell growth promoting amount of calcium ions; an effective cell growth promoting amount of glucose; an effective amount of insulin to aid cells in glucose uptake; an effective cell growth promoting amount of hydrocortisone; an effective amount of epidermal growth factor to bind epidermal growth factor receptors on cells; an effective amount of transferrin to increase DNA synthesis in cells; an effective amount of cholera toxin to increase DNA synthesis in cells; an effective amount of triiodothyronine to increase DNA synthesis in cells; and an effective growth promoting amount of mammalian hormones and mitogenic factors, including lipoprotein, cholesterol, phospholipids and fatty acids.

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Dayton, MD, United States of America

Katharine H Cole