Method for immortalizing chicken cells
AssigneeRegents of the University of Minnesota
InventorsDouglas N. Foster, James A. Farris, Linda K. Foster
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Abstract
This invention relates to the introduction of p53 under the control of the metallothionein promoter into primary cells to produce immortalized cell lines. The cells are useful as substrates for viral propagation, as contaminant-free sources for recombinant protein production, for recombinant virus production and as cell substrates to support primary cells and improve virus yield during virus propagation.
Claims14
A method for transforming chicken cells comprising the steps of:
- positioning nucleic acid encoding p53 under the control of a metallothionein promoter in a gene vector capable of directing expression of p53;
- introducing the gene vector into primary chicken cells;
- and
- selecting foci of cells with population doubling times of about 0.6 to about 1.5 population doublings per day, wherein the cells are reverse-transcriptase negative and are non-tumorigenic.
A method for propagating virus comprising the steps of:
- contacting at least one infectious virus particle with at least one cell of an immortalized chicken cell culture, wherein the cells of the culture contain a gene vector expressing p53 under the control of the metallothionein promoter;
- and
- collecting virus produced by the cells.
A method for propagating virus comprising the steps of:
- contacting at least one infectious virus particle with a primary cell;
- growing the primary cells with immortalized chicken cells containing a gene vector expressing p53 under the control of the metallothionein promoter in cell culture;
- and
- collecting virus produced from the cell culture.
Immortalized, non-transformed chicken cells containing p53 under the control of the metallothionein promoter and containing at least one vector capable of directing expression of recombinant protein in the cells.
Dependencies are inferred from the claim text.