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Updated: Jun 25, 2026
Isolation of CD133+ Liver Stem Cells for Clonal Expansion
Published on: October 10, 2011
The isolation and in vitro expansion of hepatic Sca-1 progenitor cells
Elizabeth Clayton1, Stuart J Forbes
1MRC/Centre for Inflammation Research, The Queen's Medical Research Institute, University of Edinburgh, UK. Elizabeth.Clayton@ed.ac.uk
Insights
Liver progenitor cells expand during injury. Sca1(+) cells, identified in the liver and gallbladder, show different proliferation rates on laminin versus collagen I, crucial for long-term culture.
Area of Science:
- Hepatology
- Stem Cell Biology
- Regenerative Medicine
Background:
- Liver injury triggers the expansion of intra-hepatic progenitor cells when hepatocyte proliferation is suppressed.
- Purified progenitor cells, obtained via density gradient centrifugation, contain diverse hematopoietic, epithelial, and endothelial lineages, suggesting the presence of liver stem cells.
- The precise identity of liver stem cells is still under investigation, with some evidence pointing to Sca1(+) CD45(-) cells exhibiting progenitor characteristics.
Purpose of the Study:
- To identify and characterize Sca1(+) cells in the liver and gallbladder following injury.
- To investigate the in vitro expansion and culture requirements of Sca1(+) cells.
- To determine the influence of extracellular matrix components on the proliferation of purified Sca1(+) cells versus heterogeneous progenitor populations.
Main Methods:
- Identification of Sca1(+) cells in intra-hepatic and gall bladder tissues post-liver injury.
- Ex vivo expansion of Sca1(+) cells, both as purified populations and within heterogeneous cultures.
- Assessment of Sca1(+) cell proliferation on different extracellular matrices (laminin and collagen I).
Main Results:
- Sca1(+) cells were identified in both intra-hepatic and gall bladder tissues after liver injury.
- Purified Sca1(+) cells exhibited significantly different proliferation rates when cultured on laminin compared to collagen I.
- The proliferation of heterogeneous progenitor cell populations was not affected by the tested extracellular matrices.
Conclusions:
- Laminin appears to be a critical component for the long-term culture and proliferation of Sca1(+) liver progenitor cells.
- Culture conditions, specifically the extracellular matrix, are essential for maintaining the proliferative capacity of purified Sca1(+) cells.
- These findings have implications for optimizing cell culture techniques for liver progenitor cells in regenerative medicine applications.
Abstract:
The intra-hepatic population of liver progenitor cells expands during liver injury when hepatocyte proliferation is inhibited. These cells can be purified by density gradient centrifugation and cultured. Separated by size only this population contains small cells of hematopoietic, epithelial and endothelial lineages and is thought to contain liver stem cells. The identity of liver stem cells remains unknown although there is some evidence that tissue Sca1(+) CD45(-) cells display progenitor cell characteristics. We identified both intra-hepatic and gall bladder Sca1(+) cells following liver injury and expanded ex vivo Sca1 cells as part of heterogenous cell culture or as a purified population. We found significant difference between the proliferation of Sca-1 cells when plated on laminin or collagen I while proliferation of heterogenous population was not affected by the extracellular matrix indicating the necessity for culture of Sca1(+) cells with laminin matrix or laminin producing cells in long term liver progenitor cell cultures.
