Permanent human hepatocyte cell line and its use in a liver assist device (LAD)
AssigneeBaylor College of Medicine
InventorJames H. Kelly
Life of this patent
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Abstract
Permanent cell lines are provided which display liver-specific biological activity at a level effective to support a subject suffering from hepatic failure or insufficiency. The cells find use in treating various liver disorders. An example of the cell lines is HepG2/C3A deposited under ATCC No. CRL-10741. Methods for use of the cells in liver assist devices and methods for producing liver assist devices are also provided.
Claims24
A permanent human hepatocyte cell line constitutively having liver-specific biological activity at a level effective to support a subject suffering from hepatic failure or insufficiency, wherein said cell line is capable of growing in glucose-free media, and wherein at confluence, said cell line exhibits a rate of expression of albumin of at least about 20 .mu.g/ml total cell protein/24 hours, and wherein said cell line produces albumin and alphafetoprotein such that the albumin to alphafetoprotein ratio is at least about 15.
In a liver assist device containing cells for supporting a subject suffering from hepatic failure or insufficiency, the improvement comprising cells of a permanent human hepatocyte cell line constitutively having liver-specific biological activity at a level effective to support a subject suffering from hepatic failure or insufficiency, wherein said cell line is capable of growing in glucose-free media, and wherein at confluence, said cell line exhibits a rate of expression of albumin of at least about 20 .mu.g/ml total cell protein/24 hours, and wherein said cell line produces albumin and alphafetoprotein such that the albumin to alphafetoprotein ratio is at least about 15.
A method for preparing a liver assist device having a membrane for supporting a cell line for supporting a subject suffering from hepatic failure or insufficiency comprising the steps of:
- (a) providing cells of a permanent human hepatocyte cell line constitutively having liver-specific biological activity at a level effective to support a subject suffering from hepatic failure or insufficiency, wherein said cell line is capable of growing in glucose-free media, and wherein at confluence, said cell line exhibits a rate of expression of albumin of at least about 20 .mu.g/mg total cell protein/24 hours, and wherein said cell line produces albumin and alphafetoprotein such that the albumin to alphafetoprotein ratio is at least about 15, on a semipermeable membrane having opposed first and second surfaces;
- and
- (b) culturing said cells on said first surface until the cells reach confluence, said culturing step comprising the steps of:
- (i) seeding said first surface with said cells;
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- (ii) continuously circulating culture medium which supports the growth of said cells until the cells reach confluence.
A method for producing serum proteins comprising the steps of:
- (a) maintaining a permanent hepatocyte cell line constitutively having liver-specific biological activity, wherein said cell line is capable of growing in glucose-free media, and wherein at confluence, said cell line exhibits a rate of expression of albumin of at least about 20 .mu.g/mg total cell protein/24 hours, and wherein said cell line produces albumin and alphafetoprotein such that the albumin to alphafetoprotein ratio is at least about 15, in a nutrient culture medium;
- (b) recovering supernatant fluid from said culture;
- (c) separating plasma proteins from said supernatant fluid;
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- (d) purifying said plasma proteins.
Dependencies are inferred from the claim text.