Label-Free Isolation and Single Cell Biophysical Phenotyping Analysis of Primary Cardiomyocytes Using Inertial Microfluidics

Hossein Tavassoli; Prunella Rorimpandey; Young Chan Kang; Michael Carnell; Chris Brownlee; John E Pimanda; Peggy P Y Chan; Vashe Chandrakanthan

doi:10.1002/smll.202006176

Label-Free Isolation and Single Cell Biophysical Phenotyping Analysis of Primary Cardiomyocytes Using Inertial Microfluidics

Small. 2021 Feb;17(8):e2006176. doi: 10.1002/smll.202006176. Epub 2020 Dec 28.

Authors

Hossein Tavassoli^{1

2

3}, Prunella Rorimpandey^{2

3}, Young Chan Kang^{2

3}, Michael Carnell⁴, Chris Brownlee⁵, John E Pimanda^{2

3

6

7}, Peggy P Y Chan¹, Vashe Chandrakanthan^{2

3}

Affiliations

¹ Department of Telecommunications, Electrical, Robotics and Biomedical Engineering, Swinburne University of Technology, Hawthorn, Victoria, 3122, Australia.
² Department of Pathology, School of Medical Sciences, University of New South Wales, Sydney, NSW, 2052, Australia.
³ Adult Cancer Program, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, 2052, Australia.
⁴ Biomedical Imaging Facility, Mark Wainwright Analytical Centre, University of New South Wales, Sydney, NSW, 2052, Australia.
⁵ Flow Cytometry Facility, Mark Wainwright Analytical Centre, University of New South Wales, Sydney, NSW, 2052, Australia.
⁶ Prince of Wales Clinical School, University of New South Wales, Sydney, NSW, 2052, Australia.
⁷ Department of Haematology, Prince of Wales Hospital, Sydney, NSW, 2052, Australia.

PMID: 33369875
DOI: 10.1002/smll.202006176

Abstract

To advance the understanding of cardiomyocyte (CM) identity and function, appropriate tools to isolate pure primary CMs are needed. A label-free method to purify viable CMs from mouse neonatal hearts is developed using a simple particle size-based inertial microfluidics biochip achieving purities of over 90%. Purified CMs are viable and retained their identity and function as depicted by the expression of cardiac-specific markers and contractility. The physico-mechanical properties of sorted cells are evaluated using downstream real-time deformability cytometry. CMs exhibited different physico-mechanical properties when compared with non-CMs. Taken together, this CM isolation and phenotyping method could serve as a valuable tool to progress the understanding of CM identity and function, and ultimately benefit cell therapy and diagnostic applications.

Keywords: cardiomyocyte; cell separation; deformability cytometry; microfluidics; regeneration.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Biophysics
Mice
Microfluidics*
Myocytes, Cardiac*
Single-Cell Analysis