# US 4,393,133 A · Human hepatoma derived cell line, process for preparation thereof, and uses therefor

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- Number: US 4,393,133 A
- Document ID: US4393133A
- Status: Granted
- Kind: A
- Application number: 06/158,685
- Filed: 1980-06-12
- Granted: 1983-07-12
- Earliest claimed: 1980-06-12
- Assignees: The Wistar Institute of Anatomy and Biology
- Inventors: Barbara B. Knowles; David P. Aden
- Web page: https://patentgrep.com/patent/US4393133A

## CPC

- C07K 14/005: from viruses
- C12N 5/067: {Hepatocytes}
- C12N 5/0693: {Tumour cells; Cancer cells}
- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53)
- C12N 2502/13: connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
- C12N 2730/10122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- Y10S 435/948: using viruses or cell lines
- Y10S 436/82: Hepatitis associated antigens and antibodies
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- 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)
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## Abstract

Human hepatoma cell lines, useful for metabolic studies such as screening potential carcinogens and mutagens, for cultivation of viruses, and for preparation of vaccines is obtained by culturing human hepatocarcinoma or hepatoblastoma on lethally irradiated cell feeder layers in the presence of a culture medium.

## Claims

### Claim 1

A process for assessing the metabolic conversion of chemicals and drugs considered to be potential carcinogens and mutagens, which comprises:
(a) maintaining a culture of a human hepatic cell line selected from the group consisting of the cell lines designated Hep 3B and Hep G2 in a nutrient medium,
(b) exposing said cell line to the chemical or drug to be tested,
(c) analyzing said culture for the presence of metabolites of said chemical or drug, and
(d) introducing said metabolites to cultures of other mammalian cells to determine their mutagenic capabilities.

### Claim 2 (depends on 1)

The process according to claim 1 in which said cell line comprises Hep G2.

### Claim 3 (depends on 1)

The process according to claim 1 in which said cell line comprises Hep 3B.

### Claim 4

A process for isolation of hepatitis B virus surface antigen for use as a vaccine which comprises:
(a) maintaining a human hepatic cell line, designated Hep 3B, which contains the hepatitis B virus genome in nutrient culture medium,
(b) recovering the supernatant fluid from said culture, and
(c) purifying the hepatitis B virus surface antigen in said supernatant fluid for use as a vaccine.

### Claim 5

A process for isolation of human plasma proteins which comprises:
(a) maintaining a human hepatic cell line which synthesizes plasma proteins selected from the group consisting of the cell lines designated Hep 3B and Hep G2 in a nutrient culture medium,
(b) recovering the supernatant fluid from said culture, and
(c) separating plasma proteins from said supernatant fluid, and purifying said plasma proteins.

### Claim 6 (depends on 5)

The process according to claim 5 in which said cell line comprises Hep G2.

### Claim 7 (depends on 5)

The process according to claim 5 in which said cell line comprises Hep 3B.

### Claim 8

The cell line having the identifying characteristics of Hep 3B.

### Claim 9

The cell line having the identifying characteristics of Hep G2.