Derivation and characterization of new cell line from intestine of turbot (Scophthalmus maximus)

Abstract

A continuous intestine cell line from turbot (Scophthalmus maximus) designated as SMI was established utilizing the tissue explant technique. Primary SMI cell was cultured at 24 °C in a medium with 20% fetal bovine serum (FBS), then subcultured in 10% FBS after 10 passages. Impacts of medium or temperature on the growth of SMI were examined and the results indicated it grew well in DMEM supplemented with 10% FBS at 24 °C. The SMI cell line was subcultured more than 60 times. Karyotyping, chromosome number, and ribosomal RNA genotyping analysis revealed that SMI had a modal diploid chromosome number of 44 and originated from turbot. After being transfected with pEGFP-N1 and FAM-siRNA, a large number of green fluorescence signals were observed in SMI, indicating that SMI could be used as an ideal platform to explore gene function in vitro. In addition, the expression of epithelium-associated genes such as itga6, itgb4, gja1, claudin1, zo-1, and E-cadherin in SMI suggested the SMI had some characteristics of epidermal cells. The upregulation of immune-associated genes such as TNF-β, NF-κB, and IL-1β in SMI after stimulation with pathogen-associated molecular patterns suggested the SMI might exhibit immune functions similar to the intestinal epithelium in vivo.

The manipulation of cell suspensions from zebrafish intestinal mucosa contributes to understanding enteritis

“…However, the isolation of GALT's immune cells has been performed in several cultured fish species, such as rainbow trout (Oncorhynchus mykiss) (25), carp and turbot (Scophthalmus maximus) (26). In order to analyze the proportion of immune cells in the gut, the preparation of single cells is a crucial part of flow cytometric analysis of the gut (27).…”