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A Co-culture Method to Investigate the Crosstalk Between X-ray Irradiated Caco-2 Cells and PBMC
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
A Co-culture Method to Investigate the Crosstalk Between X-ray Irradiated Caco-2 Cells and PBMC

A Co-culture Method to Investigate the Crosstalk Between X-ray Irradiated Caco-2 Cells and PBMC

DOI:
10.3791/56908-v

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11:40 min

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January 30, 2018

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, , , , , , ,
1Dipartimento di Fisica,Università degli Studi di Pavia, 2IRCCS Istituti Clinici Scientifici Maugeri

Chapters

  • 00:04Title
  • 01:05Cell Preparation and Irradiation
  • 03:21Cell Metabolic Activity (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide [MTT]) and Viability Assays, and Trans-Epithelial Electrical Resistance (TEER)
  • 06:12Western Blot Analysis
  • 09:15Results: Representative Effects of Caco-2 Cell Radiation Exposure and PBMC Co-culture
  • 11:01Conclusion

Summary

Automatic Translation

Automatic Translation

We present a protocol to investigate the crosstalk between X-ray-irradiated Caco-2 and peripheral blood mononuclear cells (PBMC). The protocol starts with Caco-2 irradiation and set-up of the co-culture with PBMC; subsequently, trans-epithelial electrical resistance is measured regularly over 48 h and western blot performed in both Caco-2 and PBMC.

Tags

Keywords: Caco-2 CellsPBMCsCocultureIonizing RadiationEpithelial Cell PermeabilityTight Junction FunctionRadiation BiologyRadioimmunotherapyCell CultureDensity Gradient CentrifugationX-ray IrradiationDosimetryCell Viability

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