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3D sheep rumen epithelial structures driven from single cells in vitro.

Zebang Xu1,2, Xinxin Xu1,2, Bin Yang1,3

  • 1Institute of Dairy Science, College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China.

Veterinary Research
|November 10, 2023
PubMed
Summary

Researchers developed a novel method to culture rumen epithelial organoids from single cells. This new in vitro system for studying ruminant digestion may reduce the need for animal testing.

Keywords:
3D cultureRNA-seqRumen epitheliumorganoid

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Area of Science:

  • Animal Science
  • Cell Biology
  • Digestive Physiology

Background:

  • Ruminants are vital livestock providing high-quality protein.
  • Existing 3D organoid models exist for abomasum and intestines, but not the rumen.
  • The rumen is crucial for lignocellulose digestion via its microbiota.

Purpose of the Study:

  • To establish a method for culturing rumen epithelial organoids from single cells.
  • To characterize the properties and potential applications of these organoids.

Main Methods:

  • Single rumen epithelial cells were cultured to form self-organized 3D structures.
  • Specific growth factors (EGF, Noggin, Wnt3a, IGF-1, FGF-10) and small molecules (CHIR-99021, A83-01, SB202190, Y-27632) were used.
  • Organoid stability was tested through passage, cryopreservation, and resuscitation.

Main Results:

  • Rumen epithelial organoids self-organized into structures resembling native rumen epithelium.
  • Key growth factors and small molecules significantly improved organoid seeding efficiency, formation, and maintenance.
  • Organoids retained functionality after cryopreservation and passage, expressing specific genes and uptaking fatty acids.

Conclusions:

  • A feasible method for culturing rumen epithelial organoids from single cells was established.
  • This novel in vitro system offers a promising alternative for studying rumen function and may reduce experimental animal use.