Method for selecting a permissive cell line for replicating avian viruses

Patent metadata

Field	Value
Publication number	US-10053672-B2
Application number	US-201414905485-A
Country	US
Kind code	B2
Filing date	Jul 24, 2014
Priority date	Jul 25, 2013
Publication date	Aug 21, 2018
Grant date	Aug 21, 2018

Abstract

The present invention relates to a method for obtaining an untransformed avian cell line enabling in vitro avian virus replication. Said method includes the following steps: a) culturing avian embryonic stem cells in the presence of a stroma for at least 3 days; b) culturing for at least 2 days in a medium having a low serum concentration; c) culturing for at least 2 days in a medium having a low serum concentration containing 1 to 10 mM of hexamethyleme bisacetamide (HMBA); d) culturing for at least 10 days in a medium having a low serum concentration; and e) culturing or freezing an avian cell line enabling avian virus replication. The invention also relates to the resulting cell line and to the use thereof in vaccine preparations.

First claim

The invention claimed is:

1. A vaccinal preparation comprising a non-transformed avian cell line allowing replication of avian viruses in vitro;
wherein the non-transformed avian cell line is made by a method comprising:
a) cultivating avian embryonic stem cells in the presence of a stroma for at least 3 days;
b) cultivating in a medium with a low serum concentration for at least 2 days;
c) cultivating in a low serum concentration medium comprising between 1and 10 mM of hexamethylene bisacetamide (HMBA), for at least 2 days; and
d) cultivating in a low serum concentration medium for at least 10 days;
to thereby provide a non-transformed avian cell line allowing replication of avian viruses,
wherein the non-transformed avian cell line expresses the following markers: CD44+, beta1 integrin, collagen 1 and OLFM3; and
wherein the non-transformed avian cell line is infected with at least one avian virus.

Assignees

• Agronomique Inst Nat Rech

Inventors

• Vautherot Jean-François
• Pain Bertrand
• Denesvre Caroline
• Fragnet-Trapp Laetitia

Classifications

• C12N5/0606

  Pluripotent embryonic cells, e.g. embryonic stem cells [ES] (embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696) · CPC title

• C12N5/0629

  Keratinocytes; Whole skin · CPC title

• C12N5/0652

  Cells of skeletal and connective tissues; Mesenchyme · CPC title

• C12N2500/30

  Organic components · CPC title

• C12N2720/10052

  relating to complementing cells and packaging systems for producing virus or viral particles · CPC title