3 results about "Bovine kidney" patented technology

This invention relates to the NBLS (Niobium Bovine Kidney Cell Line), with accession number CCTCC NO: C2022197. This invention also provides its application in culturing BVDV virus. This invention successfully introduced the SV40-LT gene into primary Niobium Bovine Kidney Cells, activating their telomerase activity and extending the cell lifespan in vitro. Experiments showed that the NBLS Niobium Bovine Kidney Cell Line of this invention can be passaged for more than 50 generations without changing its growth and proliferation characteristics, thus obtaining an immortalized cell line. This F50 generation Niobium Bovine Kidney Cell Line NBLS is highly sensitive to BVDV virus and can efficiently proliferate the virus. The virus titer proliferated using this cell line shows increased sensitivity compared to primary Niobium Bovine Kidney Cells, reducing production costs. Therefore, this invention provides a new technical means for the large-scale production of BVDV vaccines.

The present invention discloses the application of the carboxypeptidase 6 (CPA6) gene in regulating host resistance to bovine enterovirus (BEV). Using CRISPR / Cas9 gene editing technology, a bovine kidney cell line with CPA6 gene knockout and a cell model resistant to BEV virus infection were constructed. The results of virological detection showed that the proliferation rate of BEV in gene-knockout cells was significantly lower than that in wild-type cells, while complementing the CPA6 gene caused the antiviral effect of the cells to disappear, indicating that the CPA6 gene plays an important role in regulating host resistance to BEV infection. The present invention identified the function of the CPA6 gene at the cellular level and discovered for the first time that the gene can be used as a potential target for gene-edited animal design in disease-resistant breeding, prevention and treatment of BEV infection, which is of great significance for reducing drug abuse and avoiding epidemics.

The invention provides a method for separating and identifying Holstein calf renal cortex primary cells, which belongs to the technical field of cells and comprises the following steps: cleaning Holstein calf renal cortex tissue blocks twice with D-Hanks, cutting the tissue blocks to about 1mm < 3 >, adding pancreatin / EDTA (Ethylene Diamine Tetraacetic Acid), pre-digesting in a constant-temperature water tank at 37 DEG C, naturally precipitating, and discarding supernate; adding type II collagenase for resuspension, digesting in a constant-temperature water tank at 37 DEG C, collecting supernate, repeating the digesting for 7-8 times, centrifuging the supernate, adding a culture medium containing fetal calf serum, terminating the digesting, centrifuging, collecting cell precipitate, resuspending, and inoculating into a culture dish; adding a culture medium to resuspend the tissue fragments, and inoculating the resuspended tissue fragments into a new culture dish to obtain bovine renal cortex primary cells; the bovine renal cortex primary cells are identified by an immunofluorescence method, and more than 95% of the bovine renal cortex primary cells are renal cortex fibroblasts. The invention provides a method for efficiently obtaining high-activity bovine renal cortex primary cells, which is simple to operate.