TY - JOUR T1 - Development of a modular automated system for maintenance and differentiation of adherent human pluripotent stem cells JF - bioRxiv DO - 10.1101/079335 SP - 079335 AU - Duncan E. Crombie AU - Maciej Daniszewski AU - Helena H. Liang AU - Tejal Kulkarni AU - Fan Li AU - Grace E. Lidgerwood AU - Alison Conquest AU - Damian Hernández AU - Sandy S. Hung AU - Katherine P. Gill AU - Elisabeth De Smit AU - Lisa S. Kearns AU - Linda Clarke AU - Valentin M. Sluch AU - Xitiz Chamling AU - Donald J. Zack AU - Raymond C.B. Wong AU - Alex W. Hewitt AU - Alice Pébay Y1 - 2016/01/01 UR - http://biorxiv.org/content/early/2016/10/05/079335.abstract N2 - Patient-specific induced pluripotent stem cells (iPSCs) have tremendous potential for development of regenerative medicine, disease modelling and drug discovery. However, the processes of reprogramming, maintenance and differentiation are labour intensive and subject to inter-technician variability. To address these issues, we established and optimised 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. ER -