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Biologia
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Methods to Study Epithelial Transport Protein Function and Expression in Native Intestine and Caco-2 Cells Grown in 3D
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Biologia
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JoVE Journal Biologia
Methods to Study Epithelial Transport Protein Function and Expression in Native Intestine and Caco-2 Cells Grown in 3D

Methods to Study Epithelial Transport Protein Function and Expression in Native Intestine and Caco-2 Cells Grown in 3D

DOI:
10.3791/55304-v

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11:27 min

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March 16, 2017

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, , , , , , , ,
1Department of Medicine,University of Illinois at Chicago, 2Department of Research,Jesse Brown VA Medical Center

Capítulos

  • 00:05Título
  • 01:533D Caco-2 Cell Culture on a Gelatinous Protein Solution
  • 03:23Immunofluorescence Staining of Epithelial Transporters in 3D Caco-2 Cells
  • 05:12Measurement of 5-HT Uptake in Mouse Ileum
  • 06:29Equilibration and Tissue Treatment
  • 07:41Quantification of the Mucosal to serosal Flux (Jm-s) and 3[H]-5-HT Accumulation
  • 08:42Results: The Expression and Function of SERT Protein Measured in Caco-2 3D Cell Cultures and Ileal Mucosa Ex Vivo
  • 09:57Conclusion

Summary

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We describe simple methods to study the regulation of intestinal serotonin transporter (SERT) function and expression using an in vitro cell culture model of Caco-2 cells grown in 3D and an ex vivo model of mouse intestines. These methods are applicable to the study of other epithelial transporters.

Tags

Epithelial TransportSerotonin TransporterSERTCaco-2 Cells3D Cell CultureUssing ChamberIntestinal EpitheliumElectrolyte TransportersCell-cell InteractionsCell-extracellular Matrix InteractionsTransport FunctionTherapeutic Screening

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