| abstract |
A replication procedure of a virus selected from the group consisting of adenovirus, hepadnavirus, herpes virus, orthomyxovirus, papovavirus, paramyxovirus, picornavirus, poxvirus, reovirus and retrovirus, in stem cells derived from avian embryos, in which said cells are established by a procedure comprising the following steps: A) isolation, culture and expansion of avian embryonic stem cells in a complete culture medium containing all the factors that allow their growth and in the presence of an inactivated feeder layer of mouse fibroblasts and supplemented with animal serum; B) subculture by modification of the culture medium to obtain a progressive or total withdrawal of said factors, said serum and said feeder layer; C) establishment of adherent or non-adherent avian cell lines capable of proliferating in a basal environment in the absence of exogenous growth factors, inactivated feeder layer and a level of 2% or less of serum or without serum, said replication process comprising the steps of: a) proliferation of said cells obtained in stage C) in a culture vessel, in suspension, in a serum-free medium No. 1, in which said cells are allowed to aggregate with each other to form cell clusters during at least said proliferation stage, and in which said cell clusters are not broken by centrifugation and said cells are subcultured by dilution in a larger culture vessel; b) infection of said cells in cell clusters with the virus selected when the cell density is at least 1.5 million cells / ml; c) shortly before infection, simultaneously with infection, or shortly after infection, addition to the cell culture of serum-free medium No. 2; and d) further culture of said infected cells in order to allow virus replication; and e) optionally, collecting said virus. |