Method of producing an undifferentiated avian cell culture using avian primordial germ cells

摘要

A method for producing a sustained culture of undifferentiated avian cells expressing an embryonic stem cell phenotype comprises: a) collecting avian gonadal cells comprising primordial germ cells from an avian embryo after formation of the primitive streak; b) depositing the avian gonadal cells in contact with a preconditioned feeder matrix; c) growing the avian gonadal cells on the preconditioned feeder matrix in the presence of media for a time sufficient to produce an avian cell culture consisting essentially of undifferentiated avian cell expressing an embryonic stem cell phenotype. The avian embryo can be selected from a chicken, turkey, duck, goose, quail and pheasant embryo. For the production of a sustained culture of undifferentiated chicken cells expressing an embryonic stem cell phenotype, gonadal cells are collected from a stage 27 to a stage 30 chicken embryo. The gonadal cells are deposited in contact with a preconditioned mouse fibroblast feeder cell layer and grown on the cell layer in the presence of a conditioned media for a time sufficient to produce a sustained avian cell culture. The sustained avian cell culture consists essentially of undifferentiated avian cells expressing an embryonic stem cell phenotype.