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Development of a modular automated system for maintenance and differentiation of adherent human pluripotent stem cells

Citation

Crombie, DE and Daniszewski, M and Liang, HH and Kulkarni, T and Li, F and Lidgerwood, GE and Conquest, A and Hernandez, D and Hung, SS and Gill, KP and De Smit, E and Kearns, LS and Clarke, L and Sluch, VM and Chamling, X and Zack, DJ and Wong, RCB and Hewitt, AW and Pebay, A, Development of a modular automated system for maintenance and differentiation of adherent human pluripotent stem cells, SLAS Discovery, 22, (8) pp. 1016-1025. ISSN 2472-5552 (2017) [Refereed Article]


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Copyright Statement

Copyright 2017 Society for Laboratory Automation and Screening

DOI: doi:10.1177/2472555217696797

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 MultiMACS24 Separator. Pluripotency markers were assessed to confirm pluripotency of the generated iPSCs. Passaging was performed using an enzyme-free dissociation method. Proof of concept of differentiation was obtained by differentiating human PSCs into cells of the retinal lineage. Key advantages of this automated approach are the ability to increase sample size, reduce variability during reprogramming or differentiation, and enable medium- to high-throughput analysis of human PSCs and derivatives. These techniques will become increasingly important with the emergence of clinical trials using stem cells.

Item Details

Item Type:Refereed Article
Keywords:automated cell culture platform, automation, human pluripotent stem cells, iPSCs, liquid handling, passaging, retinal cell differentiation
Research Division:Medical and Health Sciences
Research Group:Ophthalmology and Optometry
Research Field:Ophthalmology
Objective Division:Health
Objective Group:Clinical Health (Organs, Diseases and Abnormal Conditions)
Objective Field:Hearing, Vision, Speech and Their Disorders
Author:Li, F (Dr Fan Li)
Author:Hewitt, AW (Dr Alex Hewitt)
ID Code:120439
Year Published:2017
Deposited By:Menzies Institute for Medical Research
Deposited On:2017-08-24
Last Modified:2017-11-22
Downloads:0

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