Isolation and Characterization of Hepatic Stem Cells, or “Oval Cells,” from Rat Livers
The pace of research on the potential therapeutic uses of liver stem cells or “oval cells” has accelerated significantly in recent years. Concurrent advancements in techniques for the isolation and characterization of these cells have helped fuel this research. Several models now exist for the induction of oval cell proliferation in rodents. Protocols for the isolation and culture of these cells have evolved to the point that they may be set up in any laboratory equipped for cell culture. The advent of magnetic cell sorting has eliminated reliance on expensive flow cytometric sorting equipment to generate highly enriched populations of oval cells. Our laboratory has had much success in using the oval cell surface marker Thy-1 in combination with magnetic sorting to produce material suitable for testing the influence of a myriad of chemical signaling molecules on the oval cell phenotype. This chapter will describe our basic strategy for oval cell induction and isolation. Additionally, two in vitro procedures are described which the reader may find useful in the early stages of developing an oval cell research project.
Key wordsLiver stem cells oval cells 2-acetylaminofluorene (2AAF) partial hepatectomy (PH) collagenase perfusion simple gravity enrichment immunomagnetic sorting (IMS) flow cytometric sorting (FACS) proliferation assay migration assay
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