Journal
/
Biology
/
Methods to Study Epithelial Transport Protein Function and Expression in Native Intestine and Caco-2 Cells Grown in 3D
JoVE Journal
Biology
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biology
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

•

11:27 min

•

March 16, 2017

•

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

Chapters

  • 00:05Title
  • 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

Automatic Translation

Automatic Translation

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

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

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/55304?article_type=v

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid
check_url/55304?article_type=v

Primary Human Bronchial Epithelial Cells Grown from Explants
check_url/55304?article_type=v

Hi-C: A Method to Study the Three-dimensional Architecture of Genomes.
check_url/55304?article_type=v

Chromatin Isolation by RNA Purification (ChIRP)
check_url/55304?article_type=v

Rapid Genetic Analysis of Epithelial-Mesenchymal Signaling During Hair Regeneration
check_url/55304?article_type=v

Induction and Analysis of Epithelial to Mesenchymal Transition
check_url/55304?article_type=v

High Resolution Whole Mount In Situ Hybridization within Zebrafish Embryos to Study Gene Expression and Function
check_url/55304?article_type=v

siRNA Screening to Identify Ubiquitin and Ubiquitin-like System Regulators of Biological Pathways in Cultured Mammalian Cells
check_url/55304?article_type=v

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein
check_url/55304?article_type=v

Using Caco-2 Cells to Study Lipid Transport by the Intestine

Read Article