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Metoder til at studere Epithelial Transport Protein Funktion og Expression i Native tarmen og Caco-2 celler dyrket i 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

Metoder til at studere Epithelial Transport Protein Funktion og Expression i Native tarmen og Caco-2 celler dyrket i 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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Vi beskriver enkle metoder til at studere reguleringen af intestinal serotonin transporter (SERT) funktion og ekspression anvendelse af en in vitro cellekultur-model af Caco-2-celler dyrkes i 3D og en ex vivo model for muse tarmene. Disse metoder kan anvendes til studiet af andre epitel transportører.

Tags

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

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