DUCK EMBRYONIC DERIVED STEM CELL LINES FOR THE PRODUCTION OF VIRAL VACCINES
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.
1 .- 23 . (canceled)
24 . A duck cell line derived from duck embryonic stem cells characterized in that it is continuous;
diploid; and
free of replication-competent endogenous retrovirus particles.
25 . The duck cell line according to claim 24 , wherein said duck is a Pekin duck.
26 . The duck cell line according to claim 24 , wherein said duck is a Muscovy duck.
27 . The duck cell line according to claim 24 , wherein said duck cell line further has at least one of the following characteristics:
a high nucleo-cytoplasmic ratio,
an endogenous telomerase activity,
a population doubling time of around 30 hours or less at 37° C., and
expresses one or more additional markers selected from the group consisting of alkaline phosphatase, SSEA-1, EMA-1, and ENS-1.
28 . The duck cell line according to claim 24 , wherein cells of said cell line are capable of proliferating in a basal medium in the absence of exogenous growth factors, a feeder layer and animal serum for at least 50 generations, at least 150 generations, or at least 250 generations.
29 . The duck cell line according to claim 24 , wherein the genome of said duck cell line does not contain proviral sequence of avian leucosis virus (ALV-E) and/or endogenous avian virus (EAV) susceptible to produce replication competent endogenous retroviral particles.
30 . The duck cell line according to claim 24 , wherein said duck cell line does not produce replication-competent endogenous avian leucosis virus (ALV-E) and/or endogenous avian virus (EAV) retrovirus particles.
31 . The duck cell line according to claim 24 , wherein said duck cell line is capable of being infected with and producing a virus in culture.
32 . The duck cell line according to claim 24 , wherein said duck cell line further comprises a nucleic acid sequence coding a virus of interest.
33 . The duck cell line according to claim 31 , wherein said virus is selected from the group comprising poxviruses, orthomyxoviruses, paramyxoviruses, herpes viruses, hepadnaviruses, adenoviruses, parvoviruses, reoviruses, circoviruses, coronaviruses, flaviviruses, togaviruses, birnavriruses and retroviruses.
34 . The continuous diploid duck cell line according to claim 31 , wherein said virus is selected from the group consisting of measles virus, Respiratory Syncytial Virus (RSV), mumps virus, rubella virus, Sendai virus, human para-influenza types I and III, Newcastle disease virus, Rinderpest virus, canine distemper virus, duck para-influenza virus, measles virus, Newcastle Disease virus (NDV), Sinbis virus, Semliki forest virus, O′nyong′nyong virus, Chikungunya virus, Mayaro virus, Ross river virus, Eastern Equine encephalitis virus, Western Equine encephalitis virus, Venezuelan Equine encephalitis virus, Marek Disease virus (MDV), duck hepatitis B virus (DHMV), heron hepatitis B virus (HHBV), snow goose hepatitis B virus (SGHBV), Bursal Disease virus, Dengue virus, Japanese encephalitis virus (JEV), West Nile virus (WNV), Infectious Bronchitis virus, Chicken Anenia virus, reticulo-endotheliosis virus, duck infectious anemia virus, suck spleen necrosis virus, duck parvovirus, fowl adenovirus, goose adenovirus, duck adenovirus, pigeon adenovirus.
35 . The continuous diploid duck cell line according to claim 31 , wherein said virus is measles virus.