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US FDA/National Center for Toxicological Research

2 ARTICLES PUBLISHED IN JoVE

Biology

Isolation of CD133+ Liver Stem Cells for Clonal Expansion

C. Bart Rountree¹, Wei Ding¹, Hein Dang¹, Colleen VanKirk², Gay M. Crooks³

¹Department of Pediatrics and Pharmacology, Pennsylvania State College of Medicine, ²Department of Pharmacology, Pennsylvania State College of Medicine, ³Department of Pediatrics, University of California Los Angeles, School of Medicine

Here we describe the isolation of CD133 expressing liver stem cells and cancer stem cells from whole murine liver, a process that requires tissue digestion, cell enrichment, and flow cytometry isolation. We include methods for advanced single cell isolation and clonal expansion.

Biology

In Vivo Alkaline Comet Assay and Enzyme-modified Alkaline Comet Assay for Measuring DNA Strand Breaks and Oxidative DNA Damage in Rat Liver

Wei Ding¹, Michelle E. Bishop¹, Lascelles E. Lyn-Cook¹, Kelly J. Davis², Mugimane G. Manjanatha¹

¹Division of Genetic and Molecular Toxicology, US FDA/National Center for Toxicological Research, ²Toxicologic Pathology Associates, US FDA/National Center for Toxicological Research

The alkaline comet assay measures DNA strand breaks in eukaryotic cells. By adding an Endonuclease III or human 8-oxoguanine-DNA-N-glycosylase digestion step, the assay can efficiently detect oxidative DNA damage. We describe methods for using these assays to detect DNA damage in rat liver.