Investigation Of Suspension And Broad-spectrum Of Animal Vaccine Cell Lines

Cells are grown in large bioreactors for scale-up virus cultivation which is harvested to produce the corresponding vaccine.At present,the level of domestic cell lines development is behind foreign countries,which hinders the development of new vaccines and the improvement of vaccine production efficiency.This paper aims at solving two key problems in the development of animal vaccine production cell lines.In other words,suspension culturing and broadening the sensitivity spectrum of viruses.On the one hand,we explore the key genes which affect cell suspension growth,on the other hand we make virus infect cells that are originally not sensitive to this virus by using genetic engineering approaches such as RNA-Seq,gene overexpression vector construction and siRNA.We have laid the foundation for establishing the platform for cell lines capable of being cultivated using suspension medium and producing multiple vaccine.The main results are summarized as below:(1)After analyzing the transcriptional data of BHK-21 cells and CHO-K1 cells,we have found 18 significant differentially expressed genes might affecting cell suspension growth and correlated with mRNA transcriptional and translation.We have constructed a network of 18 genes by GeneMANIA and studied the effect of the most critical gene PABPC1 on cell suspension growth.It was found that BHK-21 anchorage-dependent cells could quickly be adapted to the cell suspension culture after inhibiting PABPC1.(2)We have constructed the eukaryotic expression vector to express canine distemper viral receptor NECTIN4 and screened the stable expressing cell line.It was found that expressing viral receptors helps virus infect cells that are originally not sensitive to this virus and increase the variety of culturing virus in a cell line.