Development and characterization of a stable bovine intestinal sub-epithelial myofibroblast cell line from ileum of a young calf

Uprety, Tirth; Spurlin, Brionna B.; Antony, Linto; Sreenivasan, Chithra; Young, Alan J.; Li, Feng; Hildreth, Michael B.; Kaushik, Radhey S.

doi:10.1007/s11626-019-00365-0

Cited by 5 publications

(1 citation statement)

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“…Therefore, the SV40 large T antigen is often applied for genome integration resulting in cell immortalization. So far, a series of cells from different origins have been successfully immortalized into cell lines using SV40 large T antigen through all kinds of deliveries (Sun et al 2020;Uprety et al 2019;Mitani et al 2019).…”

Section: Introductionmentioning

confidence: 99%

Establishment and characterization of immortalized human vocal fold fibroblast cell lines

Chu

Fang

et al. 2023

Biotechnol Lett

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Purpose Vocal fold scarring is abnormal scar tissue in the lamina propria layer of the vocal fold. To facilitate investigation of vocal fold scarring, we established and characterized immortalized human vocal fold broblast (iHVFF) cell lines.Methods Human vocal fold broblasts were immortalized by introducing Simian virus 40 large T antigen (SV40TAg) by transfection. Successfully transfected broblasts were sorted using ow cytometry.Immuno uorescence cytochemistry and western blot were applied to analyze the expression of bronectin, vimentin, alpha-smooth muscle actin (α-SMA) and broblast activation protein (FAP). Cell proliferation rate was measured by CCK-8 assay. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to analyze the mRNA expression level. ResultsThe iHVFFs continued to proliferate for more than 30 generations and appeared spindle-shaped.The expression of Vimentin and α-SMA were detected in both iHVFFs and primary broblasts, and enhanced expression of FAP was observed in iHVFFs. Furthermore, iHVFFs exhibited an increased proliferative capability compared with the primary broblasts. RT-qPCR results suggested that collagen type III alpha 1 chain (COL3A1), interleukin-6, cyclooxygenase 2 (COX2), hyaluronan synthase 2 (HAS2), hepatocyte growth factor (HGF) in the iHVFFs signi cantly increased, whereas transforming growth factor-β1 (TGF-β1), elastin and matrix metallopeptidase-1 (MMP-1) expression signi cantly downregulated. No differences in mRNA expression of α-SMA, bronectin and collagen type I alpha 2 chain (COL1A2) were noted between iHVFFs and primary broblasts.Conclusion iHVFFs can be used as a novel tool cell for future researches on the mechanisms of pathogenesis and treatment of vocal fold scarring.

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Section: Introductionmentioning

confidence: 99%