Compositions and methods useful for HCV infection
AssigneeVertex Pharmaceuticals Incorporated
InventorsAnn Kwong, Randal Byrn, Lola M Reid
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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.
Claims15
A composition comprising:
- a. 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 up to 1 year after birth;
- b. a feeder cell, wherein the feeder cell is capable of providing an extracellular matrix and diffusible factors;
- and
- c. a Hepatitis C virus (“HCV”),
- wherein the HCV is RNA 898 (“RNA 898” deposited on Mar. 27, 2001, in the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110-2209;
- ATCC Deposit No.:
- PTA-3237).
A cell mixture comprising liver cells and hematopoietic cells, the cells being released from a liver of a human aged three months or older after conception up to 1 year after birth, wherein:
- each component of the cell mixture has a size that allows it to pass through a 40-micron filter;
- and
- the cell mixture also comprises cells that express alpha fetoprotein, cells that express albumin, and cells that express glycophorin, but is free of detectable cells that express CD34 protein.
A composition comprising a cell mixture, serum-free media, and an HCV, the composition being prepared by the following steps:
- a. dissecting a liver of a human aged three months or older after conception up to 1 year after birth in a buffer comprising ethylene glycol bis(β-aminoethyl ether)-N,N,N′,N′-tetraacctate (EGTA);
- b. incubating the dissected liver in a buffer comprising collagenase to separate cells from the liver;
- c. passing the separated cells through a 40-micron filter;
- d. removing red blood cells from the filtered cells;
- c. resuspending the cells of step (d) in scrum-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 or any combination of them;
- f. culturing the cells in the serum-free media of step (e);
- and
- g. infecting the cells of step (f) with an HCV.
A composition comprising:
- a. 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 up to 1 year after birth, wherein each component of the cell mixture has a size that allows it to pass through a 40-micron filter;
- b. an extracellular matrix;
- and
- c. an HCV, wherein the HCV is RNA 898.
A composition comprising:
- a. 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 up to 1 year after birth, wherein each component of the cell mixture has a size that allows it to pass through a 40-micron filter;
- b. a feeder cell, wherein the feeder cell is capable of providing an extracellular matrix and diffusible factors;
- and
- c. an HCV, wherein the HCV is RNA 898.
Dependencies are inferred from the claim text.