Pennsylvania State College of Medicine

3 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.

Medicine

Standardized Histomorphometric Evaluation of Osteoarthritis in a Surgical Mouse Model

William J. Pinamont¹, Natalie K. Yoshioka¹, Gregory M. Young¹, Vengadeshprabhu Karuppagounder¹, Elijah L. Carlson¹, Adeel Ahmad¹, Reyad Elbarbary^1,2, Fadia Kamal^1,3

¹Center for Orthopedic Research and Translational Sciences, Department of Orthopedics and Rehabilitation, Pennsylvania State College of Medicine, ²Department of Biochemistry and Molecular Biology, Pennsylvania State College of Medicine, ³Department of Pharmacology, Pennsylvania State College of Medicine

The current protocol establishes a rigorous and reproducible method for quantification of morphological joint changes that accompany osteoarthritis. Application of this protocol can be valuable in monitoring disease progression and evaluating therapeutic interventions in osteoarthritis.