Victor Chang Cardiac Research Institute

3 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.