Cytotechnology

, Volume 68, Issue 4, pp 713–724 | Cite as

X/XO or H2O2 induced IPEC-J2 cell as a new in vitro model for studying apoptosis in post-weaning piglets

  • Xuan Cai
  • Lihui Zhu
  • Xiaolian Chen
  • Yongshuai Sheng
  • Qi Guo
  • Jian Bao
  • Jianxiong Xu
Original Research

Abstract

We previously demonstrated that intestinal epithelial cell apoptosis in weaned piglets is much more serious than that observed in sucking piglets and is related to oxidative stress during weaning. It is difficult to study the apoptosis mechanisms only using in vivo methods because of the limit of existing research technology. An in vitro cellular system is required for piglet intestinal epithelial cell apoptosis research. In this study, a non-tumorigenic epithelial cell line, IPEC-J2 cells, was employed as a cell model. Hydrogen peroxide and xanthine/xanthine oxidase (X/XO) were both used and compared for apoptosis modeling. The concentrations of hydrogen peroxide and XO were selected and verified using cell viability analysis, the comet assay and flow cytometry. Intracellular ROS were measured using fluorescent probes. Additionally, the expression levels of the apoptosis-related genes Fas, Bcl-2, P53, Caspase 3, Caspase 8, and Caspase 9 were analyzed using quantitative RT-PCR. The results indicated the optimal modeling method is a final concentration of 0.5 mM H2O2 incubated with IPEC-J2 cells for 1 h at 37 °C in 5 % CO2 for hydrogen peroxide-induced apoptosis modeling, and a final concentration of 250 μM X/50 U/L XO incubated with IPEC-J2 cells for 6 h at 37 °C in 5 % CO2 for X/XO-induced apoptosis modeling. For the apoptotic pathway, the X/XO modeling method is more similar to 21 days weaning piglets. Therefore, we suggest that X/XO modeling with IPEC-J2 cells be used as an in vitro cell culture model for weaning piglet intestinal epithelial cell apoptosis.

Keywords

Apoptosis Oxidative stress Cell model IPEC-J2 Weaning 

Supplementary material

10616_2014_9823_MOESM1_ESM.doc (552 kb)
Supplementary material 1 (DOC 553 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Xuan Cai
    • 1
    • 2
  • Lihui Zhu
    • 1
    • 2
  • Xiaolian Chen
    • 3
  • Yongshuai Sheng
    • 1
    • 2
  • Qi Guo
    • 1
    • 2
  • Jian Bao
    • 1
    • 2
  • Jianxiong Xu
    • 1
    • 2
  1. 1.School of Agriculture and BiologyShanghai Jiao Tong UniversityShanghaiChina
  2. 2.Shanghai Key Laboratory for Veterinary and BiotechnologyShanghaiChina
  3. 3.Institute of Animal Husbandry and Veterinary ScienceJiangxi Academy of Agricultural SciencesNanchangChina

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