Journal article

Development of a Modular Automated System for Maintenance and Differentiation of Adherent Human Pluripotent Stem Cells

DE Crombie, M Daniszewski, HH Liang, T Kulkarni, F Li, GE Lidgerwood, A Conquest, D Hernández, SS Hung, KP Gill, E De Smit, LS Kearns, L Clarke, VM Sluch, X Chamling, DJ Zack, RCB Wong, AW Hewitt, A Pébay

Slas Discovery | SAGE PUBLICATIONS INC | Published : 2017

Abstract

Patient-specific induced pluripotent stem cells (iPSCs) have tremendous potential for development of regenerative medicine, disease modeling, and drug discovery. However, the processes of reprogramming, maintenance, and differentiation are labor intensive and subject to intertechnician variability. To address these issues, we established and optimized protocols to allow for the automated maintenance of reprogrammed somatic cells into iPSCs to enable the large-scale culture and passaging of human pluripotent stem cells (PSCs) using a customized TECAN Freedom EVO. Generation of iPSCs was performed offline by nucleofection followed by selection of TRA-1-60–positive cells using a Miltenyi MultiM..

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Grants

Awarded by Robert and Joan Dircks Foundation


Funding Acknowledgements

The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by grants from the Ophthalmic Research Institute of Australia (10.13039/501100001108), the Joan and Peter Clemenger Foundation, the Philip Neal bequest, a National Health and Medical Research Council Practitioner Fellowship (10.13039/501100000925 to A.W.H.), an Australian Research Council Future Fellowship (10.13039/501100000923 FT140100047 to A.P., FT140100047), the University of Melbourne and Operational Infrastructure Support from the Victorian Government.