hTERT-BTY cell line牛甲状腺永生化细胞株细胞系 BioVector NTCC质粒载体菌种细胞基因保藏中心

hTERT-BTY cell line牛甲状腺永生化细胞株细胞系 BioVector NTCC质粒载体菌种细胞基因保藏中心
The stable bovine thyroid (BTY) cell line hTERT-BTY from primary BTY cells was established by telomerase reverse transcriptase over expression. The results of karyotype analysis and experiments on morphological and biological characteristics indicated that this cell line possessed the qualities of primary BTY cells, which could be extended indefinitely with stable morphology and steady growth rates. The hTERT-BTY cell line, has 60 chromosomes including 29 pairs of autosomes and 1 pair of sex chromosomes without structure aberrations. It can express thyroid-specific function genes thyroid-stimulating hormone receptor and sodium/iodide symporter in high abundance ratios. The cell line is sensitive to FMDV strains and is expected to be used as a powerful tool for FMDV clinical diagnosis, separation, detection and culture. The different mRNA expression levels in infected and uninfected hTERT-BTY cells were analyzed to identify the pathways of immunity using RNA-seq. The results suggested that the hTERT-BTY cell line could be regarded as an effective tool for the immune response exploration of FMDV.Primary BTY cells were isolated from BTY tissues to establish an immortalized BTY cell line. Previous studies confirmed that the shortening of telomeres was the reason for cell senescence in most normal somatic cells. Therefore, the over-expression of the telomerase reverse transcriptase (TERT) gene and activation of telomerase are critical strategies in cellular immortality. The hTERT expression plasmid was transduced into primary BTY cells and the G418-resistant colonies were isolated. One colony was expanded and cultured, this colony was named hTERT-BTY. Confluent monolayers of the hTERT-BTY cells at passages 10–100 complied with typical primary BTY cell morphology was shown in Figure 1. Growth curves of hTERT-BTY cells at passages 5, 10, 100, and 220 were created (Figure 1). The cell line showed a steady growth rate in passages 5–220, whereas the viability of the primary BTY cells began to decline after passage 6. Taken together, the results suggested that the hTERT-BTY cell line could be extended indefinitely with steady morphology and growth rate.  DMEM/F12 mediumsupplemented with 200 μg/mL G418, 10% (v/v) fetal calf serum, 10 μg/mL insulin, 5 μg/mL transferrin, 4 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin
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