Newborn sertoli-cell immortalization cell line and establishing method and application thereof

Abstract

The invention discloses a newborn sertoli-cell immortalization cell line and an establishing method and application thereof. The newborn sertoli-cell immortalization cell line is named as the newbornsertoli-cell immortalization cell line hTERT-NBSC, and the collection number of the hTERT-NBSC is CCTCC NO:C2018201. Tests find that the hTERT-NBSC can continuously pass 65 generations or above, and the growth and proliferation characteristic is kept constant. The cell line is sensitive to inoculation of ORFV, the ORFV virus can be efficiently proliferated, and the uniformity and the stability ofthe virus can be guaranteed. The titer of the ORFV virus proliferated by the cell line is equivalent to that of the newborn sertoli cell, the cell sorting rate of the ORFV virus is higher than that ofthe NBSC, and the cell sorting rate can be 1:3. Meanwhile, vaccine is prepared with the passage cell line, the production technology is simplified, the production cycle is shortened, the production cost is reduced, and the stable quality of the contagious ovine ecthyma vaccine prepared with the method is also guaranteed. Therefore, through the newborn sertoli-cell immortalization cell line and the establishing method and application thereof, a new technological mean is provided for large-scale production of the contagious ovine ecthyma vaccine.

Description

technical field [0001] The invention relates to an immortalized cell line and its establishment method and application, in particular to an immortalized bovine testis Sertoli cell line, its establishment method and its application in culturing, proliferating and culturing sheep infectious impetigo virus. The invention belongs to the technical field of cell engineering. Background technique [0002] Sertoli cell (SC) is the epithelial cell on the basement membrane of the seminiferous tubule and the only somatic cell in the seminiferous tubule. Structurally, it provides a scaffold for the differentiation and maturation of germ cells, provides support, nutrition and protection for germ cells, and is called the "nanny cell" of germ cells. In the process of sperm differentiation and maturation, Sertoli cells not only deliver gonadotropins to the seminiferous epithelium, control a series of dynamic processes of mitosis, meiosis and subsequent differentiation of spermatogenic cell...

AI Technical Summary

Problems solved by technology

ORFV can grow on skin cells, kidney cells of cattle and sheep, and testicular cells of calves and lambs, and produce cytopathic changes, but the virus titer is low. Ovine infectious impetigo virus has the advantages of convenient cell culture, high virus isolation efficiency, and high virus proliferation titer, but there are problems in obtaining the cells (sheep embryos are required), and a large number of animals are required to provide tissues in vaccine production.

[0007] In China, although the Veterinary Research Institute of Qinghai Academy of Animal Husbandry and Veterinary Sciences and the Gansu Provincial Institute of Animal Husbandry and Veterinary Medicine have developed an attenuated vaccine for impetigo contagiousis, but the preparation process is complicated and backward, and newborn cattle must be used to breed the attenuated vaccine for impetigo contagiosum. primary testis cells

However, the procedures for preparing this kind of cells are complicated, the production cost is high, and the production efficiency is low, which brings inconvenience to the large-scale production of sheep infectious impetigo attenuated vaccine by vaccine factories, and it is difficult for enterprises to obtain huge profits.

At the same time, the variety of newborn bovine testicles used for each breeding of sheep aphthous antigen is different, which makes it difficult to ensure the quality and stability of the attenuated vaccine produced. In addition, newborn bovine testicles have the potential risk of carrying other pathogens

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