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Poznan University of Medical Sciences

3 ARTICLES PUBLISHED IN JoVE

Biology

Selecting and Isolating Colonies of Human Induced Pluripotent Stem Cells Reprogrammed from Adult Fibroblasts

Urszula Polak^1,2, Calley Hirsch¹, Sherman Ku³, Joel Gottesfeld³, Sharon Y.R. Dent¹, Marek Napierala¹

¹Department of Molecular Carcinogenesis and Center for Cancer Epigenetics, University of Texas M.D. Anderson Cancer Center, ²Department of Cell Biology, Poznan University of Medical Sciences, ³Department of Molecular Biology, The Scripps Research Institute

We present a protocol for efficient reprogramming of human somatic cells into human induced pluripotent stem cells (hiPSC) using retroviral vectors encoding Oct3/4, Sox2, Klf4 and c-myc (OSKM) and identification of correctly reprogrammed hiPSC by live staining with Tra-1-81 antibody.

Biology

Methods to Classify Cytoplasmic Foci as Mammalian Stress Granules

Anaïs Aulas^*1,2, Marta M. Fay^*1,2, Witold Szaflarski^1,2,3, Nancy Kedersha^1,2, Paul Anderson^1,2, Pavel Ivanov^1,2,4

¹Division of Rheumatology, Immunology, and Allergy, Brigham and Women's Hospital, ²Department of Medicine, Harvard Medical School, ³Department of Histology and Embryology, Poznan University of Medical Sciences, ⁴The Broad Institute of Harvard and M.I.T.

Stress Granules (SGs) are nonmembranous cytoplasmic structures that form in cells exposed to a variety of stresses. SGs contain mRNAs, RNA-binding proteins, small ribosomal subunits, translation-related factors, and various cell signaling proteins. This protocol describes a workflow that uses several experimental approaches to detect, characterize, and quantify bona fide SGs.

Cancer Research

Detection of Lung Tumor Progression in Mice by Ultrasound Imaging

Nour Ghaddar^1,2, Shuo Wang¹, Véronique Michaud¹, Urszula Kazimierczak^1,3, Nicolas Ah-son¹, Antonis E. Koromilas^1,4

¹Lady Davis Institute for Medical Research, Sir Mortimer B. Davis-Jewish General Hospital, ²Division of Experimental Medicine, Faculty of Medicine, McGill University, ³Department of Cancer Immunology, Chair of Medical Biotechnology, Poznan University of Medical Sciences, ⁴Gerald Bronfman Department of Oncology, Faculty of Medicine, McGill University

This protocol describes the steps taken to induce KRAS lung tumors in mice as well as the quantification of formed tumors by ultrasound imaging. Small tumors are visualized in early timepoints as B-lines. At later timepoints, relative tumor volume measurements are achieved by the measurement tool in the ultrasound software.