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Cell Symposia | 10 Years of iPSCs

September 25, 2016 - September 27, 2016

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CDI Poster Presentations

Development and Functional Applications of Human iPSC-derived Spinal Motor Neurons

J Liu1, C Chavez1, B Meline1, KH Kim1, A Essex2, E Jones1, C McMahon1, WB Wang1

1). Cellular Dynamics International, A FUJIFILM Company, Madison, WI
2). PhenovistaBiosciences, San Diego, CA

Abstract

The aim of this study was to produce spinal motor neurons from human iPSCswith sufficient purity for use in a multitude of downstream assays. In particular we wanted to produce motor neurons that could be cultured in defined conditions over long periods of time, without being hampered by outgrowth from proliferative cell types. Using an optimized differentiation protocol that improves upon published methods, we were able to produce motor neurons from iPSCsat greater than 60% purity as measured by Isl1/2 and Tuj1 positive staining. These cells can be stored frozen, thawed, and cultured in media without glia for extended periods. We collected ICC, qPCR and MEA data to characterize the motor neuron cells and used iPSC lines from multiple donors to demonstrate a robust protocol that produces motor neurons independent of donor iPSC line. These data show the characteristics and utilization of motor neurons produced from iPSC.

Excitatory Cortical Neurons (iCell GlutaNeurons) Derived from Human iPS Cells Create Functional Macro Networks In Vitro

C Kannemeier, E Enghofer,L Harms, L Norkosky, R Lewis, B Swanson
Cellular Dynamics International, A FUJIFILM Company, Madison, WI

Abstract

The ability to produce human neuronal populations from iPSCs combined with advancements in micro electrode array instrumentation make it now possible to study human neuronal network activity in vitro. This poster presents data demonstrating the functional neuronal network properties of iCell GlutaNeurons, a human iPSC-derived excitatory cortical neuron population that enables electrophysiology and excitatory toxicity assays. Using single cell gene expression as a guide, we established a robust differentiation process starting from iPS cells that generates primarily cortical glutamatergic neurons. iCell GlutaNeurons react to increasing amounts of glutamic acid with increased cell death exhibiting excitatory toxicity. Pre-treatment of iCell GlutaNeurons with NMDA and AMPA receptor inhibitors AP5 and DNQX inhibited excitatory toxicity. Most importantly, the cells show a robust formation of a synaptically driven macro network with spontaneous, synchronous electrical activity in the MEA platform. The synchronous activity can be reversibly inhibited by AP5 and DNQX, thus demonstrating the possibility to modulate iCell GlutaNeurons electrophysiological activity using pharmacology.

First, Large-scale, Patient-derived iPSC Initiative with iPSCs Banked from Over 1700 Donors in 2 Years and Counting…

A Mack, WB Wang, C Kannemeier, E Nuwaysir, A Bosch, M Collins, T Novak
Cellular Dynamics International, A FUJIFILM Company, Madison, WI

Abstract

We demonstrate the production of an unprecedented number of quality controlled iPSCs using Cellular Dynamics International’s (CDI) non-­integrating, plasmid‐based reprogramming platform. This CIRM initiative supports iPSC manufacture from up to 3000 donors spanning multigenic disorders within 3 years. Product uniformity has been sought through automation, sample batching, and a custom Laboratory Information Management System (LIMS). CDI has successfully reprogrammed and banked the requisite number of clones from over 72% (1745) of the 2426 donor samples received and around 60% (1424) of the donor samples received have been successfully banked for distribution to meet and even exceed all grant milestones to date. Furthermore, the potential of these clones to form multiple germ layers was validated by injecting lines generated from the first 25 donor samples reprogrammed into SCID mice. CDI’s robust process results in a bank of consistently derived iPSCs less subject to variability and more likely to improve the downstream data collected from their use.

Details

Start:
September 25, 2016
End:
September 27, 2016
Website:
http://www.cell-symposia-ipscs.com/

Organizer

Cell Press

Venue

Claremont Hotel & Spa
41 Tunnel Road
Berkeley, CA 94705 United States
Website:
http://www.fairmont.com/claremont-berkeley/