Accedi

University of Technology, Sydney

5 ARTICLES PUBLISHED IN JoVE

Biology

Isolation of Small Noncoding RNAs from Human Serum

Samantha Khoury¹, Pamela Ajuyah¹, Nham Tran^2,3

¹School of Medical and Molecular Biosciences, Faculty of Science, University of Technology, Sydney, ²Centre for Health Technologies, Faculty of Engineering and Information Technology, University of Technology, Sydney, ³The Sydney Head and Neck Cancer Institute, Sydney Cancer Centre, Royal Prince Alfred Hospital

This protocol describes a method for extracting small RNAs from human serum. We have used this method to isolate microRNAs from cancer serum for use in DNA arrays and also singleplex quantitative PCR. The protocol utilizes phenol and guanidinium thiocyanate reagents with modifications to yield high quality RNA.

Biology

Super-resolution Imaging of the Cytokinetic Z Ring in Live Bacteria Using Fast 3D-Structured Illumination Microscopy (f3D-SIM)

Lynne Turnbull^*1, Michael P. Strauss^*1, Andrew T. F. Liew¹, Leigh G. Monahan¹, Cynthia B. Whitchurch¹, Elizabeth J. Harry¹

¹The ithree Institute, University of Technology, Sydney

Spatiotemporal information about dynamic proteins inside live cells is crucial for understanding biology. A type of super-resolution microscopy called fast 3D-structured illumination microscopy (f3D-SIM) reveals unique information about the cytokinetic Z ring in bacteria: both its bead-like appearance and the rapid dynamics of FtsZ within the ring.

Bioengineering

Layered Alginate Constructs: A Platform for Co-culture of Heterogeneous Cell Populations

Poonam Sharma¹, Julianne D. Twomey¹, Michelle Patkin¹, Adam H. Hsieh¹

¹Fischell Department of Bioengineering, University of Maryland

Engineering and analysis of load bearing tissues with heterogeneous cell populations are still a challenge. Here, we describe a method for creating bi-layered alginate hydrogel discs as a platform for co-culture of diverse cell populations within one construct.

Bioengineering

Transplantation of a 3D Bioprinted Patch in a Murine Model of Myocardial Infarction

Christopher D. Roche^1,2,3,4, Carmine Gentile^1,2,3

¹The University of Sydney, ²University of Technology Sydney (UTS), ³The Royal North Shore Hospital, ⁴University Hospital of Wales

This protocol aims to transplant a 3D bioprinted patch onto the epicardium of infarcted mice modeling heart failure. It includes details regarding anesthesia, the surgical chest opening, permanent ligation of the left anterior descending (LAD) coronary artery and application of a bioprinted patch onto the infarcted area of the heart.

Bioengineering

Cardiac Spheroids as in vitro Bioengineered Heart Tissues to Study Human Heart Pathophysiology

Poonam Sharma^1,2,3,4, Carmine Gentile^2,3,4

¹University of Newcastle, ²University of Sydney, ³Kolling Institute of Medical Research, Royal North Shore Hospital, ⁴University of Technology, Sydney

This protocol aims to fabricate 3D cardiac spheroids (CSs) by co-culturing cells in hanging drops. Collagen-embedded CSs are treated with doxorubicin (DOX, a cardiotoxic agent) at physiological concentrations to model heart failure. In vitro testing using DOX-treated CSs may be used to identify novel therapies for heart failure patients.