Establishment of a lipopolysaccharide-induced inflammation model of human fetal colon cells

Abstract

Abstract AIM: To culture a human fetal colon (FHC) cell line in vitro and establish an FHC cell inflammation model that meets the requirements for high expression of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). Methods: FHC cells were cultured with various concentrations of Escherichia coli lipopolysaccharide (LPS) in appropriate media for 0.5, 1, 2, 4, 8, 16 and 24 h to stimulate an inflammatory reaction. The viability of FHC cells was detected by a Cell Counting Kit-8 (CCK-8) assay. The transcriptional levels of IL-6 and tumor necrosis factor-α and the protein expression changes in FHC cells were detected by PCR and ELISA, respectively. Appropriate stimulation conditions were selected (i.e., LPS concentration and treatment time), based on changes in cell survival rate, and IL-6 and TNF-α expression levels. Results: An LPS concentration higher than 100 μg/mL or a treatment time longer than 24 h resulted in morphological changes and decreased cell survival. By contrast, expression levels of IL-6 and TNF-α significantly increased within 24 h when LPS concentration lower than 100 μg/mL and peaked at 2 h, whilst maintaining cell morphology and viability in FHC cells. Conclusion: The treatment of FHC cells with 100 μg/mL LPS within 24h was optimal. was optimal in terms of stimulating IL-6 and TNF-α expression.