US FDA/National Center for Toxicological Research
2 ARTICLES PUBLISHED IN JoVE
Biology
Isolation of CD133+ Liver Stem Cells for Clonal Expansion
C. Bart Rountree 1, Wei Ding 1, Hein Dang 1, Colleen VanKirk 2, Gay M. Crooks 3
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 Ding 1, Michelle E. Bishop 1, Lascelles E. Lyn-Cook 1, Kelly J. Davis 2, Mugimane G. Manjanatha 1
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.
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