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

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Abstract

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-α (TNF-α).

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-α 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-α expression levels. 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 24 h was optimal in terms of stimulating IL-6 and TNF-α expression.

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Data Availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

Not applicable.

Funding

The present study was supported by the National Natural Science Foundation (grant no. 82173360), Kuanren Talents Program of the second affiliated hospital of Chongqing Medical University (grant no. 13-003-023), Senior Medical Talents Program of Chongqing for Young and Middle-aged (grant no. 11–020), Chongqing medical scientific research project (Joint project of Chongqing Health Commission and Science and Technology Bureau) (grant no. 2020GDRC014), China Postdoctoral Science Foundation (grant no. 2022M720606), and Special support for postdoctoral of Chongqing (grant no. 2212013362080033).

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Authors and Affiliations

  1. Department of Gastroenterology, The Second Affiliated Hospital of Chongqing Medical University, 74 Linjiang Road, Yuzhong, Chongqing, 400010, P.R. China

    Keqi Yu, Shengtao Liao, Chuanfei Li, Ya Song, Zhechuan Mei & Lin Lv

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  1. Keqi Yu
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  2. Shengtao Liao
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  3. Chuanfei Li
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  4. Ya Song
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  5. Zhechuan Mei
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  6. Lin Lv
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Contributions

LL and ZM designed the study and performed the experiments, KY and YS collected the data, KY and SL analyzed the data, KY and CL prepared the manuscript. All authors read and approved the final manuscript.

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Correspondence to Zhechuan Mei or Lin Lv.

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Yu, K., Liao, S., Li, C. et al. Establishment of a lipopolysaccharide-induced inflammation model of human fetal colon cells. Mol Biol Rep 50, 5557–5564 (2023). https://doi.org/10.1007/s11033-023-08465-7

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