The United States of America As Represented by the Secretary of

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

Other

John F Lechner

Curtis C Harris

Roger R Reddel

Katharine H Cole

Johng S Rhim

Douglas E Brash

Brenda I Gerwin

Paul Amstad

Robert T Su

Andrea M Pfeifer

Brent E Korba

George H Yoakum

Yang Ke

Ke Yang

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

Immortalized human bronchial epithelial and human mesothelial cell lines have been obtained. Various uses of these cell lines have been described.

Immortalized non-tumorigenic human bronchial epithelial cell lines

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.

Cell culture medium for human liver epithelial cell line

Immortalized human bronchial epitherial mesothelial cell lines

A modified protoplast fusion method and cell line is disclosed that stably transfects human cells with pSV2-derived plasmids at frequencies greater than 10.sup.-3. This procedure makes it possible to test the biological effect of individual genes (i.e., oncogenes and other cellular genes, and viral genes). To demonstrate the utility of this invention, a pSV2gpt.sup.+ plasmid constructed to carry a subgenomic fragment of hepatitis B virus (HBV) that contained the core antigen gene (HBc gene) is transfected into human cells. Human cell lines are stably transfected with the HBC.sup.+ gene by selecting recipient cells for expression of guanine phosphoribosyl transferase expression; other selective markers, i.e., neomycin resistance, can also be used. Conditions for enhancing the expression of the transfected gene(s) have also been developed. For example, with this gpt.sup.+ /HBc.sup.+ cell line it is shown that growth in serum-free medium or treatment with 5'-azacytidine stimulates the production of the HBV core antigen.

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Bethesda, MD, United States of America

John F Lechner