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Establishment and Genetic Manipulation of Murine Hepatocyte Organoids
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Biologie
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Journal JoVE Biologie
Establishment and Genetic Manipulation of Murine Hepatocyte Organoids

Establishment and Genetic Manipulation of Murine Hepatocyte Organoids

DOI:
10.3791/62438-v

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14:54 min

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February 12, 2022

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, , ,
1Key Laboratory of Experimental Teratology, Ministry of Education, Institute of Molecular Medicine and Genetics, School of Basic Medical Sciences, Stem Cell and Regenerative Medicine Research Center, Cheeloo College of Medicine,Shandong University, 2The Research Center of Stem Cell and Regenerative Medicine,Shandong University Cheeloo Medical College, School of Basic Medical Sciences, Shandong University

Chapitres

  • 00:01Introduction
  • 00:47Murine Hepatocyte Isolation by Liver Perfusion and Digestion
  • 02:47Hepatocyte Organoids Culture
  • 04:29Single Cells from Hepatocyte Organoids
  • 07:22siRNA or Plasmid Transfection
  • 12:58Representative Results
  • 13:58Conclusion

Summary

Traduction automatique

Traduction automatique

A long-term ex vitro 3D organoid culture system was established from mouse hepatocytes. These organoids can be passaged and genetically manipulated by lentivirus infection of shRNA/ectopic construction, siRNA transfection and CRISPR-Cas9 engineering.

Tags

Keywords: HepatocyteOrganoidsLiverMetabolismDetoxificationRegenerationIn Vitro CultureGenetic ModificationLiver PerfusionLiver DigestionSingle Cell SuspensionOrganoid CultureMetrogelPassageGenetic Manipulation

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