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Related Experiment Videos

An efficient method of sorting liver stem cells by using immuno-magnetic microbeads.

Yu-Fei He1, Yin-Kun Liu, Dong-Mei Gao

  • 1Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai 200032, China.

World Journal of Gastroenterology
|May 24, 2006
PubMed
Summary

A new method efficiently isolates highly pure fetal mouse liver stem cells using immuno-magnetic microbeads. This technique provides a fast, viable, and cost-effective tool for stem cell research.

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Area of Science:

  • Stem Cell Biology
  • Hepatology
  • Cell Sorting Technologies

Background:

  • Liver stem cells are crucial for liver regeneration and disease modeling.
  • Efficient isolation of viable liver stem cells is essential for research.
  • Current methods can be time-consuming and yield low purity.

Purpose of the Study:

  • To develop a rapid and efficient method for isolating liver stem cells.
  • To achieve high purity and viability in the isolated cell population.

Main Methods:

  • Fetal mouse liver cells were analyzed for surface antigens c-Kit and CD45/TER119 via flow cytometry.
  • Immuno-magnetic microbeads were employed for cell sorting.
  • Identification and characterization involved clone-forming assays, RT-PCR, and immunofluorescence.

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Main Results:

  • A c-Kit-(CD45/TER119)- cell population was purified with 97.9% purity using immuno-magnetic microbeads in a single step.
  • The isolation yield was approximately 6% of total sorted cells, with over 98% cell viability.
  • Cultured cells demonstrated high self-renewal and clone-forming capacity, expressing hepatocyte and bile duct cell markers.

Conclusions:

  • The developed method enables fast, efficient, and cost-effective enrichment of high-purity, viable liver stem cells.
  • This technique serves as a valuable tool for subsequent liver stem cell research.
  • Mature hepatocytes were observed in vitro after 21 days of culture.