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Development of a modular automated system for maintenance and differentiation of adherent human pluripotent stem cells
journal contribution
posted on 2023-05-19, 10:19 authored by Crombie, DE, Daniszewski, M, Liang, HH, Kulkarni, T, Li, F, Lidgerwood, GE, Conquest, A, Hernandez, D, Hung, SS, Gill, KP, De Smit, E, Kearns, LS, Clarke, L, Sluch, VM, Chamling, X, Zack, DJ, Wong, RCB, Alexander HewittAlexander Hewitt, Pebay, APatient-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.
History
Publication title
SLAS DiscoveryVolume
22Issue
8Pagination
1016-1025ISSN
2472-5552Department/School
Menzies Institute for Medical ResearchPublisher
Sage Publications, Inc.Place of publication
United StatesRights statement
Copyright 2017 Society for Laboratory Automation and ScreeningRepository Status
- Restricted