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Murdoch University

5 ARTICLES PUBLISHED IN JoVE

Biochemistry

Purification of Biotinylated Cell Surface Proteins from Rhipicephalus microplus Epithelial Gut Cells

Thomas P. Karbanowicz¹, Ala Lew-Tabor^1,2, Manuel Rodriguez Valle¹

¹Queensland Alliance for Agriculture & Food Innovation, The University of Queensland, ²Centre for Comparative Genomics, Murdoch University

A modified density centrifugation gradient-based methodology was utilized to isolate epithelial cells from Rhipicephalus microplus gut tissue. Surface-bound proteins were biotinylated and purified through streptavidin magnetic beads for utilization in downstream applications.

Biology

The Murine Choline-Deficient, Ethionine-Supplemented (CDE) Diet Model of Chronic Liver Injury

Jully Gogoi-Tiwari¹, Julia Köhn-Gaone¹, Corey Giles², Dirk Schmidt-Arras³, Francis D. Gratte^1,4, Caryn L. Elsegood¹, Geoffrey W. McCaughan^5,6,7, Grant A. Ramm^8,9, John K. Olynyk^10,11, Janina E.E. Tirnitz-Parker^1,12

¹School of Biomedical Sciences & Curtin Health Innovation Research Institute, Curtin University, ²School of Public Health & Curtin Health Innovation Research Institute, Curtin University, ³Institute of Biochemistry, Christian-Albrechts-University, ⁴School of Veterinary and Life Sciences, Murdoch University, ⁵Centenary Institute of Cancer Medicine and Cell Biology, The University of Sydney, ⁶Royal Prince Alfred Hospital, ⁷A.W. Morrow Gastroenterology and Liver Centre, ⁸QIMR Berghofer Medical Research Institute, ⁹Faculty of Medicine and Biomedical Sciences, The University of Queensland, ¹⁰Fiona Stanley and Fremantle Hospitals, ¹¹School of Medical and Health Sciences, Edith Cowan University, ¹²School of Medicine and Pharmacology, University of Western Australia

Here we describe a common method to induce chronic liver injury in mice by feeding of a choline-deficient and ethionine-supplemented (CDE) diet. We demonstrate health monitoring, liver perfusion, isolation, and preservation. A time course of six weeks can inform about liver injury, pathohistology, fibrosis, inflammatory, and liver progenitor cell responses.

Biochemistry

Untargeted Liquid Chromatography-Mass Spectrometry-Based Metabolomics Analysis of Wheat Grain

Hayley Abbiss^1,2,3, Joel P. A. Gummer^1,4, Michael Francki^5,6, Robert D. Trengove¹

¹Research and Innovation, Murdoch University, ²Centre for Digital Agriculture, Curtin University, ³Centre for Integrative Metabolomics and Computational Biology, School of Science, Edith Cowan University, ⁴Centre for Computational and System Medicine, Health Futures Institute, Murdoch University, ⁵Department of Primary Industries and Regional Development, ⁶State Agricultural Biotechnology Centre, Murdoch University

A method for the untargeted analysis of wheat grain metabolites and lipids is presented. The protocol includes an acetonitrile metabolite extraction method and reversed phase liquid chromatography-mass spectrometry methodology, with acquisition in positive and negative electrospray ionization modes.

Medicine

A Mouse Model of Incompletely Resected Soft Tissue Sarcoma for Testing (Neo)adjuvant Therapies

Francois X. Rwandamuriye^1,2, Breana J. Weston^2,3, Terrance G. Johns², W. Joost Lesterhuis^1,2, Rachael M. Zemek²

¹School of Biomedical Sciences, University of Western Australia, ²Telethon Kids Institute, University of Western Australia, ³College of Science, Health, Engineering and Education, Murdoch University

In this protocol, we describe a mouse model of incomplete surgical resection of soft tissue sarcoma for testing (neo)adjuvant therapies.

Medicine

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

Michael A. Pearen^*1, Hong Kiat Lim^*1, Francis D. Gratte^2,3, Manuel A. Fernandez-Rojo^1,4,5, Sujeevi K. Nawaratna⁶, Geoffrey N. Gobert⁷, John K. Olynyk^8,9, Janina E. E. Tirnitz-Parker^2,10, Grant A. Ramm^1,4

¹Hepatic Fibrosis Group, QIMR Berghofer Medical Research Institute, ²School of Pharmacy and Biomedical Sciences, Curtin Health Innovation Research Institute, Curtin University, ³School of Veterinary and Life Sciences, Murdoch University, ⁴School of Medicine, The University of Queensland, ⁵Madrid Institute for Advanced Studies (IMDEA) in Food, CEI UAM+CSIC, ⁶School of Medicine, Griffith University, ⁷School of Biological Sciences, Queen's University Belfast, ⁸Department of Gastroenterology & Hepatology, Fiona Stanley and Fremantle Hospital Group, ⁹School of Medical and Health Sciences, Edith Cowan University, ¹⁰Centre for Cell Therapy and Regenerative Medicine, School of Biomedical Sciences, University of Western Australia

This protocol provides a simple and reliable method for the production of viable precision-cut liver slices from mice. The ex vivo tissue samples can be maintained under laboratory tissue culture conditions for multiple days, providing a flexible model to examine liver pathobiology.