Neonatal bovine testicular sertoli cell immortalized cell line and its establishment method and application

Abstract

The invention discloses an immortalized cell line of neonatal bovine testis Sertoli cells, its establishment method and application. The immortalized Sertoli cell line of newborn bovine testis is named as the immortalized Sertoli cell line of newborn bovine testis hTERT‑NBSC, and its preservation number is CCTCC NO: C2018201. Experiments have found that hTERT-NBSC can be continuously passaged for more than 65 passages, and the growth and proliferation characteristics remain unchanged. The cell line is more sensitive to ORFV inoculation, can multiply ORFV virus efficiently, and can ensure the uniformity and stability of the virus. The virus titer of ORFV propagated by this cell line is equivalent to that of Sertoli cells of newborn bovine testis, and the cell division rate is higher than that of NBSC, and the division rate can reach 1:3. At the same time, the passage cell line is used to prepare the vaccine, which simplifies the production process, shortens the production cycle, reduces the production cost, and ensures the stable quality of the prepared sheep contagious impetigo vaccine. Therefore, the proposal of the present invention provides a new technical means for the large-scale production of the sheep contagious impetigo 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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