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Single Cell Profiling Identifies Key Pathways Expressed by iPSCs Cultured in Different Commercial Media

39 Pages Posted: 6 Jun 2018 Publication Status: Published

See all articles by Maciej Daniszewski

Maciej Daniszewski

University of Melbourne - Department of Surgery

Quan Nguyen

University of Queensland - Institute for Molecular Bioscience

Hun Chy

Monash University - Australian Regenerative Medicine Institute (ARMI)

Vikrant Singh

University of Tasmania - School of Medicine

Duncan Crombie

University of Melbourne - Department of Surgery

Tejal Kulkarni

University of Melbourne - Department of Surgery

Helena Liang

University of Melbourne - Department of Surgery

Grace Lidgerwood

University of Melbourne - Department of Surgery

Damian Hernández

University of Melbourne - Department of Surgery

Alison Conquest

University of Melbourne - Department of Surgery

Louise Rooney

University of Melbourne - Department of Surgery

Sophie Chevalier

University of Melbourne - Department of Surgery

Stacey Andersen

University of Queensland - School of Chemistry and Molecular Biosciences

Anne Senabouth

University of Queensland - Institute for Molecular Bioscience

James Vickers

University of Tasmania - Wicking Dementia Research and Education Centre

David Mackey

University of Western Australia - Centre for Ophthalmology and Visual Science

Jamie Craig

Flinders University

Andrew Laslett

Monash University - Australian Regenerative Medicine Institute (ARMI); CSIRO Manufacturing, Research Way

Alex Hewitt

University of Tasmania

Joseph E. Powell

University of Queensland - Institute for Molecular Bioscience

Alice Pébay

University of Melbourne - Department of Surgery

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Abstract

We assessed the pluripotency of human induced pluripotent stem cells (iPSCs) maintained on an automated platform using StemFlexTM and TeSRTM-E8TM media. Analysis of transcriptome of single cells revealed similar expression of core pluripotency genes, as well as genes associated with naive and primed states of pluripotency. Analysis of individual cells from four samples consisting of two different iPSC lines each grown in the two culture media revealed a shared subpopulation structure with three main subpopulations different in pluripotency states. By implementing a machine-learning approach, we estimated that most cells within each subpopulation are very similar between all four samples. The single cell RNA sequencing analysis of iPSC lines grown in both media is the first report of the molecular signature of StemFlex medium and how it compares to that observed in TeSR-E8 medium.

Keywords: Induced Pluripotent Stem Cells; Automated Stem Cell System; Automation; Passaging; Maintenance, Pluripotency

Suggested Citation

Daniszewski, Maciej and Nguyen, Quan and Chy, Hun and Singh, Vikrant and Crombie, Duncan and Kulkarni, Tejal and Liang, Helena and Lidgerwood, Grace and Hernández, Damian and Conquest, Alison and Rooney, Louise and Chevalier, Sophie and Andersen, Stacey and Senabouth, Anne and Vickers, James and Mackey, David and Craig, Jamie and Laslett, Andrew and Hewitt, Alex and Powell, Joseph E. and Pébay, Alice, Single Cell Profiling Identifies Key Pathways Expressed by iPSCs Cultured in Different Commercial Media (2018). Available at SSRN: https://ssrn.com/abstract=3188388 or http://dx.doi.org/10.2139/ssrn.3188388
This version of the paper has not been formally peer reviewed.

Maciej Daniszewski (Contact Author)

University of Melbourne - Department of Surgery

185 Pelham Street
Carlton, Victoria 3053
Australia

Quan Nguyen

University of Queensland - Institute for Molecular Bioscience

St. Lucia
Brisbane
Australia

Hun Chy

Monash University - Australian Regenerative Medicine Institute (ARMI)

Wellington Road
Clayton, Victoria
Australia

Vikrant Singh

University of Tasmania - School of Medicine

French Street
Sandy Bay
Tasmania, 7250
Australia

Duncan Crombie

University of Melbourne - Department of Surgery

185 Pelham Street
Carlton, Victoria 3053
Australia

Tejal Kulkarni

University of Melbourne - Department of Surgery

185 Pelham Street
Carlton, Victoria 3053
Australia

Helena Liang

University of Melbourne - Department of Surgery

185 Pelham Street
Carlton, Victoria 3053
Australia

Grace Lidgerwood

University of Melbourne - Department of Surgery

185 Pelham Street
Carlton, Victoria 3053
Australia

Damian Hernández

University of Melbourne - Department of Surgery

185 Pelham Street
Carlton, Victoria 3053
Australia

Alison Conquest

University of Melbourne - Department of Surgery

185 Pelham Street
Carlton, Victoria 3053
Australia

Louise Rooney

University of Melbourne - Department of Surgery

185 Pelham Street
Carlton, Victoria 3053
Australia

Sophie Chevalier

University of Melbourne - Department of Surgery

185 Pelham Street
Carlton, Victoria 3053
Australia

Stacey Andersen

University of Queensland - School of Chemistry and Molecular Biosciences

St. Lucia
Brisbane
Australia

Anne Senabouth

University of Queensland - Institute for Molecular Bioscience

St. Lucia
Brisbane
Australia

James Vickers

University of Tasmania - Wicking Dementia Research and Education Centre

Hobart
Australia

David Mackey

University of Western Australia - Centre for Ophthalmology and Visual Science ( email )

35 Stirling Highway
Crawley, WA Western Australia 6009
Australia

Jamie Craig

Flinders University

GPO Box 2100
Adelaide S.A. 5001, SA 5063
Australia

Andrew Laslett

Monash University - Australian Regenerative Medicine Institute (ARMI)

Wellington Road
Clayton, Victoria
Australia

CSIRO Manufacturing, Research Way

Australia

Alex Hewitt

University of Tasmania ( email )

French Street
Sandy Bay
Tasmania, 7250
Australia

Joseph E. Powell

University of Queensland - Institute for Molecular Bioscience

St. Lucia
Brisbane
Australia

Alice Pébay

University of Melbourne - Department of Surgery ( email )

185 Pelham Street
Carlton, Victoria 3053
Australia