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

Background Inflammatory bowel disease (IBD) is a global health problem and there are few cell models for IBD at present. 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-a (TNF-alpha). Methods and results 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 and protein expression changes of IL-6 and TNF-a in FHC cells were detected by Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) and Enzyme-Linked Immunosorbent Assay (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-alpha expression levels. An LPS concentration higher than 100 mu g/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-alpha significantly increased within 24 h when LPS concentration lower than 100 mu g/ mL and peaked at 2 h, whilst maintaining cell morphology and viability in FHC cells. Conclusion The treatment of FHC cells with 100 mu g/mL LPS within 24 h was optimal in terms of stimulating IL-6 and TNF-alpha expression.