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- Title
中华鳖胚胎肝成纤维细胞的原代培养及其聚肌苷酸胞苷酸(Poly I:C)刺激模型构建.
- Authors
王水涛; 何盛盛; 江玲丽; 高有领
- Abstract
The aims of the present study were to establish an in vitro isolation and culture system for the embryonic liver fibroblast of Chinese soft-shelled turtle(Pelodiscus sinensis), and to study the effect of polyinosinic acid cytidylic acid (Poly I:C) stimulation on the key factors of the cellular interferon production pathway, finally, to establish the activation model of interferon production pathway based on the fibroblast. The embryo developed to stage 23 was used to obtain the liver fibroblast by trypsin digesting. After that, the cell was identified by PCR method. Its biological characteristics including cell morphology and growth pattern were analyzed. Besides, the gene and protein expression levels of the factors related to the interferon production signal pathway were detected at 24 h after cell transfection with Poly I:C. The results showed that the obtained embryonic liver fibroblasts were in good growth condition and had been successfully passed to the 20th generation. The growth curve of the cell was S-shaped, which conformed to the cell proliferation law. Fibroblast marker genes VIM and ACTA2 were confirmed. Before and after cryopreservation, the cell survival rates were 90.51% and 82.95%, respectively. After Poly I:C stimulation, the protein expressions of RIG-Ⅰ and MAVS were up-regulated, the gene expression of RIG-Ⅰ was up-regulated. Whereas, the gene expressions of m6A and YTHDF2 were down-regulated. In addition, the change of cell morphology was recorded. In conclusion, the study successfully established an in vitro isolation and culture system for embryonic liver fibroblasts and constructed an activation model of interferon production pathway based on embryonic liver fibroblasts of Chinese soft-shelled turtle.
- Publication
Acta Agriculturae Zhejiangensis, 2022, Vol 34, Issue 1, p60
- ISSN
1004-1524
- Publication type
Article
- DOI
10.3969/j.issn.1004-1524.2022.01.08