Lamb sheep sustentacular cells as well as separation method and application thereof

Abstract

The invention discloses a lamb sheep sustentacular cell as well as a separation method and application thereof, and the preservation number of the lamb sheep sustentacular cell SCST is CCTCC No.C2019203. The SCST cell disclosed by the invention can be directly used for large-scale propagation of the sheep orf virus; or transfecting a telomerase gene to immortalize the telomerase gene, then carrying out pressure coarse screening, pressure seed separation and continuous passage to obtain a subclone cell line, and the subclone cell line is applied to large-scale breeding of the orf virus. The subclone cell line can be cultured by a culture medium containing domestic serum, so that the production cost is reduced, and the subclone cell line is suitable for large-scale production and application. Compared with conventional primary cells, the subclone cell line provided by the invention has the advantages that the separation rate of the orf virus can be obviously improved, the pathogenesis time of the orf virus is obviously shortened, the copy number of the separated virus is improved by more than 100 times compared with that of the primary cells, thus providing a tool for large-scale propagation of the orf virus and research on a virus infection pathogenesis mechanism.

Description

technical field [0001] The invention relates to the technical field of cell engineering, in particular to a lamb testicular Sertoli cell, and the invention also relates to a separation method and application of the cell. Background technique [0002] Contagious Ethyma (CE), commonly known as sheep mouth sore, is an acute and contagious infectious disease caused by sheep mouth ulcer virus (ORFvirus, ORFV) to which goats, sheep, and mainly lambs are susceptible. Zoonotic diseases that can infect humans and other animals. ORFV is mainly prevalent in Inner Mongolia, Tibet, Xinjiang, Yunnan, Gansu, Sichuan, Ningxia and other sheep-intensive areas. In recent years, it has frequently occurred in Ningxia, Inner Mongolia, Xinjiang, Tibet, Shandong, Hubei, Yunnan, Shaanxi, Heilongjiang, Qinghai and other regions . The typical symptoms of cauliflower-like hyperplasia will appear on the lips, oral mucosa, tongue and nostrils of affected sheep, which seriously affects their feeding and...

AI Technical Summary

Problems solved by technology

[0003] The preparation of vaccines for the prevention of oral ulcers mainly includes the following aspects: first, through continuous separation and cultivation of primary cells of cattle or sheep, the oral oral ulcer virus is isolated from them, and then continuously passaged for attenuation to prepare attenuated vaccines; second, the current There is also a trend of research and development of inactivated vaccines; third, because the surface of the sheep oral ulcer virus is covered with a capsule shaped like a small tube, which is arranged in a criss-cross pattern to cover its epitope, resulting in less neutralizing antibodies produced in the sheep, and the protection rate for sheep In view of the particularity of this immunology, some researchers used ultrasound and other means to destroy the capsule of ORFV and then prepare a mouth sore vaccine. However, the above situations are limited to laboratory research and have not been used in production large scale use

The main reasons are: the separation of primary cells is time-consuming, laborious and expensive, the number of passages of primary cells is small, the toxicity of the cultured aphthus virus is low, the production volume within a batch is small, and the repeatability between batches is difficult to be consistent due to differences in primary cells

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