Victor Chang Cardiac Research Institute

4 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Large-Scale Production of Cardiomyocytes from Human Pluripotent Stem Cells Using a Highly Reproducible Small Molecule-Based Differentiation Protocol

Hananeh Fonoudi^*1,2,3,8, Hassan Ansari^*1,8, Saeed Abbasalizadeh¹, Gillian M Blue^6,7, Nasser Aghdami¹, David S Winlaw^6,7, Richard P Harvey^2,3,4, Alexis Bosman^*2,3, Hossein Baharvand^*1,5

¹Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, ²Developmental and Stem Cell Biology Division, Victor Chang Cardiac Research Institute, ³St. Vincent´s Clinical School, Faculty of Medicine, University of New South Wales, ⁴School of Biotechnology and Biomolecular Sciences, University of New South Wales, ⁵Department of Developmental Biology, University of Science and Culture, ⁶Heart Centre for Children, The Children´s Hospital at Westmead, ⁷Sydney Medical School, University of Sydney, ⁸Department of Developmental Biology, University of Science and Culture, Tehran, Iran

Here, we present a robust, fast and scalable cardiomyocyte differentiation protocol for human pluripotent stem cells (hPSCs). Cardiomyocytes derived using this large-scale method can provide sufficient cell numbers for their effective use in human cardiovascular disease modeling, high-throughput drug screening, and potentially clinical applications.

JoVE Journal

Rapid In Vivo Fixation and Isolation of Translational Complexes from Eukaryotic Cells

Yoshika Janapala^*1, Katrina Woodward^*1, Jiwon Lee², Melanie Rug², Thomas Preiss^1,3, Nikolay E. Shirokikh¹

¹Division of Genome Sciences and Cancer, The John Curtin School of Medical Research, The Australian National University, ²Centre for Advanced Microscopy, The Australian National University, ³Victor Chang Cardiac Research Institute

We present a technique to rapidly stabilize translational (protein biosynthesis) complexes with formaldehyde crosslinking in live yeast and mammalian cells. The approach enables dissecting transient intermediates and dynamic RNA:protein interactions. The crosslinked complexes can be used in multiple downstream applications such as in deep sequencing-based profiling methods, microscopy, and mass-spectrometry.

Biology

Anesthesia and Intubation of Preadolescent Mouse Pups for Cardiothoracic Surgery

Jianxin Wu^*1, Amy M. Nicks^1,2, Justin J. Skowno^3,4, Michael P. Feneley^1,2, Robert M. Graham^1,2, Siiri E. Iismaa^*1,2

¹Molecular Cardiology and Biophysics Division, Victor Chang Cardiac Research Institute, ²St Vincent’s Clinical School, University of New South Wales Sydney, ³Department of Anaesthesia, The Children’s Hospital at Westmead, ⁴School of Child and Adolescent Health, University of Sydney

Cardiothoracic surgical models in mice >7 days old require intubation, but this is challenging for preadolescent (8-14-day-old) mouse pups and there is little information on anesthetic regimens for intubation. Here, we present dosage regimens of ketamine/xylazine/atropine in 10-day-old C57BL/6J mouse pups that allow endotracheal intubation, while minimizing animal mortality.

Medicine

Using an Isolated Working Rat Heart Model to Test Donor Heart Preservation Strategies

Jeanette E. Villanueva^1,2, Ling Gao¹, Sanjay Dutta^1,2, Aoife Doyle¹, Yashutosh Joshi^1,2, Peter S. Macdonald^1,3,4

¹Cardiac Physiology and Transplantation, Victor Chang Cardiac Research Institute, ²St Vincent's Healthcare Clinical Campus, University of New South Wales, ³Heart and Lung Transplant Unit, St Vincent's Hospital, ⁴Faculty of Medicine, University of New South Wales

We describe an ex vivo isolated working rat heart protocol to test donor heart preservation strategies. This paper describes the protocol for use in static cold storage of rodent donor hearts; however, the protocol can also be used for donor hearts obtained after donation after circulatory death and brain death.