Compositions and methods useful for hcv infection
AssigneeVertex Pharmaceuticals Incorporated
InventorsAnn Kwong, Randal Byrn, Lola M. Reid
Life of this patent
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Abstract
The present invention provides compositions comprising cells that can effectively produce HCV after HCV infection, compositions for culturing the cells, methods for making the composition and methods for infecting the cells in the composition with HCV. The present invention also provides methods for assaying HCV production and methods for evaluating compounds that affect the production of HCV.
Claims20
1-15. (canceled)
A composition comprising a serum-free media comprising calcium, free fatty acids (FFAs), high density lipoprotein (HDL), nicotinamide, trace elements, epidermal growth factor (EGF), insulin, transferrin, hydrocortisone, and not low density lipoprotein (LDL) and, optionally, any one of the ingredients selected from the group consisting of glucagon, liver growth factor, ethanolamine and thyrotropin releasing factor.
A method for isolating and cultivating a cell mixture comprising the steps of:
- a. dissecting a liver of a human aged three months or older after conception in a buffer comprising ethylene glycol bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetate (EGTA);
- b. incubating the dissected liver in a buffer comprising collagenase to separate cells from the liver;
- c. remove objects 40 micron or greater from the separated cells;
- d. removing red blood cells from the separated cells;
- e. resuspending the cells of step (d) in a serum-free media, comprising calcium, FFAs, HDL, nicotinamide, trace elements, EGF, insulin, transferrin, hydrocortisone, and optionally, further comprising any one of the ingredients selected from the group consisting of:
- glucagon, liver growth factor, ethanolamine and thyrotropin releasing factor;
- and
- f. culturing the cells in the serum-free media of step (e).
A method for infecting cells with Hepatitis C Virus (HCV) comprising the step of contacting a composition comprising a cell mixture comprising liver cells and hematopoietic cells, the cells being released from the liver of a human aged three months or older after conception, with the HCV virus RNA898 or its infectious equivalent.
A method for assaying HCV infection comprising the steps of:
- a. incubating a composition comprising a cell mixture comprising liver cells and hematopoietic cells, the cells being released from the liver of a human aged three months or older after conception, with a feeder cell;
- b. contacting the cells in the composition with RNA898 or its infectious equivalent;
- and
- c. measuring the presence of the HCV RNA associated with the cells of the composition, the media in which the cells are cultured or both the cells and the media.
A method for evaluating the ability of a compound to affect the production of an HCV comprising the steps of:
- a. incubating a composition comprising a cell mixture comprising liver cells and hematopoietic cells, the cells being released from the liver of a human aged three months or older after conception, with a feeder cell;
- b. contacting the cells in the composition with RNA898 or its infectious equivalent;
- c. administering the compound to the composition before or after the cells are contacted with RNA898 or its infectious equivalent;
- d. measuring the HCV associated with the cells, the media in which the cells are cultured, or both the cells and the media.
Dependencies are inferred from the claim text.