Generation of duck cell lines

摘要

The present invention relates to the development and manufacturing of viral vaccines. In particular, the invention relates to the field of industrial production of viral vectors and vaccines, more in particular to the use of avian embryonic stem cells, preferably the EBx® cell line derived from duck embryonic stem cells, for the production of viral vectors and viruses. The invention is particularly useful for the industrial production of viral vaccines to prevent viral infection of humans and animals.

主权项

1. A method of making a duck cell line comprising:
a) isolating a duck embryo around oviposition; b) dissociating the embryo of step a) into cells; c) seeding the dissociated cells of step b) in basal medium comprising insulin like growth factor 1 (IGF-1), ciliary neurotrophic factor (CNTF), animal serum, and a layer of feeder cells; d) culturing the cells of step c) for at least one passage; e) withdrawing the IGF-1 and CNTF from the culture of step d); f) culturing the cells of step e) for at least one passage; g) progressively withdrawing the feeder cells from the culture of step f); h) culturing the cells of step g) for at least one passage; i) progressively withdrawing the animal serum from the culture medium of step h) over several passages; and j) adapting the cells of step i) to suspension culture conditions such that a duck cell line capable of proliferating in basal medium in the absence of exogenous growth factors, a feeder layer, and animal serum is obtained, wherein the duck cell line does not produce replication-competent endogenous retrovirus particles.