IMMORTALIZED CHICKEN EMBRYO FIBROBLASTS Russian patent published in 2019 - IPC C12N5/71 C12N7/02 C12N15/85 A61K39/12 

FIELD: biochemistry.

SUBSTANCE: group of inventions relates to the field of genetic engineering, in particular to immortalized chicken embryo fibroblasts (CEF) for virus propagation, methods for their preparation, to cell cultures, including immortalized CEF, to methods of virus replication of birds in such cells and to methods of vaccines production. Stably transfected immortalized CEF expresses the SV40 T antigen, expresses chicken telomerase (cTERT) under the control of a heterologous promoter, does not include exogenous retroviral DNA with a long terminal repeat, is capable of continuous proliferation and provides more than 45 population doubling. Methods of producing immortalized CEF include transfection of primary CEF cells using one or more DNA molecules which do not contain LTR sequences and include inverted transposon repeats, gene coding SV40 T antigen under control of suitable promoter, gene coding cTERT under control of suitable promoter, gene coding transposase under control of suitable promoter. As a result, CEF cells are selected, which are cultivated for at least 45 cell cycles.

EFFECT: produced CEF cells have a fibroblast morphology preserved during passaging, and are a substratum for replication of avian viruses or viral vector of birds for further vaccines production.

16 cl, 9 dwg, 1 ex