The ISSCR annual meeting brings together stem cell researchers from around the world to share their work, discuss tools and techniques, and advance stem cell science and regenerative medicine. The many presentations, workshops, and discussions provide opportunities to learn, collaborate, and find inspiration. You won’t want to miss the latest news and developments in stem cell science and technology.
Visit us at booth #707.
CDI Activity at ISSCR 2017
Thursday, 15 June | 11:30AM | Level 2, Room 258C
Speakers: Coby Carlson, Strategic Marketing Manager, Christopher McMahon, R&D Senior Group Leader
Cellular Dynamics International, a FUJIFILM Company
Abstract: Parkinson’s disease (PD) affects approximately 1% of people over the age of 65 and is the second most common neurodegenerative disorder after Alzheimer’s disease. The advent of induced pluripotent stem cell (iPSC) technology now grants us access to previously unattainable cell types in the human brain. In this presentation, we will discuss how stem cells are being used not only to study PD in the lab, but also to develop treatments for PD using cells as therapies. Specifically, we have created an isogenic disease model for PD using human iPSC-derived midbrain dopaminergic neurons and have developed in vitro assays for comparative analysis of mitochondrial bioenergetics, calcium handling, and network-level electrophysiology. In the regenerative medicine space, we are actively manufacturing cGMP HLA “superdonor” iPSC lines for universal utility and will provide an update on our progress in developing iPSC-derived cellular therapies, including our program to treat PD by engrafting human midbrain dopaminergic neurons.
Date: Friday, June 16 | 7:00 – 8:00 pm
Poster: F-2110 | Session III – Even
Presenting Author: Coby Carlson
Authors: Kwi Hye Kim, Kile Mangan, Sarah Dickerson, Thomas Burke, Lauren Little, Lucas Chase, Brad Swanson, Susan DeLaura, Eugenia Jones
Cellular Dynamics International – a FUJIFILM Company
Parkinson’s disease (PD) is the second most common neurodegenerative brain disorder after Alzheimer’s, affecting more than 10 million people worldwide with 60,000 Americans being newly diagnosed each year. The pathophysiological decline associated with PD is generally thought to be caused by an aberrant deterioration of dopaminergic neurons located in the substantia nigra. PD has clearly been linked to mutations occurring in several different genes including SNCA, which encodes the alpha-synuclein (a-syn) protein that is predominantly expressed at presynaptic terminals. The a-syn protein acts as the predominate aggregate within lewey bodies, a hallmark biomarker of PD. A mutation in a-syn at A53T renders the protein more susceptible to aggregation and accumulation, which has rendered this mutation to be highly penetrant and one of the most widely studied PD mutations. Interestingly, the mutant threonine (T) is actually the murine wild-type residue at this position, bringing into question the relevance of this mutation in existing PD mouse models. The advent of induced pluripotent stem cell (iPSC) technology has allowed the opportunity to study disease-specific mutations (such as A53T) in physiologically-relevant human cell cultures. Furthermore, the rise of cutting-edge genome-engineering techniques enables the generation of isogenic controls for use in phenotypic screening and disease modeling. Here we harness the power of iPSC technique to investigate the A53T mutation and its aberrant activities within human, dopaminergic neuronal cell cultures. In this poster, we present data comparing human midbrain dopaminergic neurons derived from healthy (WT) and mutant (A53T) iPS cells. We evaluate gene expression, mitochondrial bioenergetics, calcium handling, and network-level electrophysiological behaviors of both healthy and mutant cell cultures. The observed differences in these assays suggest the a-syn A53T mutant displays early pathophysiological changes tilted towards a more connected, highly-active neuronal network. In correlation with the expected pathology, current studies are ongoing to determine if “aging” cultures display the disease-relevant synaptic deterioration, including dendritic atrophy, a-syn aggregation, and / or additional mitochondrial dysfunction.
Date: Friday, June 16 | 6:00 – 7:00 pm
Poster: F-1151| Session III – Odd
Presenting Author: Eugenia Jones
Authors: Kwi Hye Kim, Kile Mangan, Susan DeLaura, Christian Kannemeier, Rachel Lewis, Michael Hancock, Brad Swanson, Coby Carlson
Cellular Dynamics International – a FUJIFILM Company
Human cell types differentiated from induced pluripotent stem cells (iPSC) offer a unique source of cellular material for toxicity screening. For example, several studies have been presented on the use of iPSC-derived cardiomyocytes and hepatocytes in safety toxicology investigations. Equally important is comparative neurotoxicity assessment in neuronal cell types for safety toxicology and uncovering molecular mechanisms underlying excitotoxic cell death pathways. Advances in iPSC technology provide access to previously unattainable cell types from the human brain opening new opportunities to address the shortcomings and limitations of rodent primary cells and immortalized cell lines Here we present the neurotoxic effects of the excitatory neurotransmitter glutamate and related compounds across a panel of cell types, including iPSC-derived GABAergic and glutamatergic cortical neurons, as well as midbrain dopaminergic neurons. For comparison, the cytotoxicity of a broad spectrum kinase inhibitor, staurosporine (STS), was also evaluated in parallel. To achieve robust signals across these different iPSC-derived neurons, we have optimized the cell culture protocols (i. e. , media, time in culture, cell plating density, etc. ). Under the various conditions tested, we observed differential responses for glutamatergic compounds (e. g. glutamate, NMDA, AMPA, and kainic acid) versus STS. Importantly, toxicity induced by glutamate could be reversed with antagonists of the AMPA and NMDA receptors, DNQX and D-AP5, respectively, suggesting the toxicity responses were due to excitotoxic effects of neuronal synaptic receptors. . We also provide measurements of electrical activities of iPSC-derived neurons on multi-electrode arrays (MEA) to assess the effects of both developmental and environmental neurotoxicants. Overall, these iPSC-derived neurons exhibit functional glutamate pathways that respond appropriately to known agonists and antagonists, thus providing biologically relevant models for identifying emerging targets for excitotoxicity research. Together with the developmental and environmental toxicity studies, these data establish a clear utility for iPSC-derived neurons in toxicology studies.
Presenting Author: Tenneille Ludwig
Authors: T. Ludwig4, A. Reimer3, T. Foroud1, D. Smith1, C. Wegel1, C. Mitchell1, M. Frasier3, A. Naito3, S. Lasch2, K. Maerk2, C. Dunifon1
- Indiana University, Indianapolis, IN
- Institute for Neurodegenerative Disorders, New Haven, CT
- Michael J. Fox Foundation, New York, NY
- WiCell – Madison, WI
The Parkinson’s Progression Markers Initiative (PPMI) is a longitudinal observational study conducted at over thirty international sites that collects data and biospecimen from idiopathic Parkinson’s patients, age-matched controls, and participants with risk factors for Parkinson’s disease (PD), such as genetic mutations, hyposmia, and REM Sleep Behavior Disorder (RBD), for up to 5 years. PPMI makes these data and biospecimen available in real time to qualified researchers to enable biomarker research. In addition to traditional biospecimen, PPMI is also committed to obtaining and distributing a range of cell lines, including uniformly collected fibroblasts and induced pluripotent stem cells (iPSCs), from these well-characterized participants to be used for biomarker research, therapeutic development, drug screening, and disease modeling.
PPMI includes fibroblast and iPSC collection as part of two separate ancillary studies. The first sub-study, performed in collaboration with the New York Stem Cell Foundation (NYSCF), derived fibroblasts and iPSCs from skin biopsies. There are fibroblasts and iPSCs from 20 idiopathic PD patients and 5 controls from one U.S. site currently available for request from this ancillary study. In order to provide cell lines to as many qualified researchers as possible, PPMI is prioritizing the expansion of these iPSC and fibroblast resources by WiCell and the Rutgers University Cell and DNA Repository (RUCDR) respectively before distributing these materials.
The second ancillary study is currently being conducted in collaboration with Cellular Dynamics International (CDI). This expanded ancillary study shifted to a blood-based collection protocol and is being carried out at 10 international sites. The sites participating in this ancillary study aim to have 135 collections from idiopathic PD patients, controls, prodromal participants, and affected and unaffected carriers of genetic mutations associated with PD by mid-2017. The iPSCs generated in collaboration with CDI are made available on a rolling basis as they complete reprogramming. All PPMI cell lines are housed at the biorepository at Indiana University. To apply for access to PPMI cell lines, please visit http://www.ppmi-info.org/access-data-specimens/request-cell-lines/.
GMP-compatible iPSC Derivation from Multiple Perinatal Tissue Sources from the Same Donors
Authors: Amanda Mack1, Katherine S. Brown2, Elizabeth Faust1, Kevin Monroe1, Ryan Wachowiak1, Wen Bo Wang1, Matthew L. Skiles2, Heather L. Brown2,
- Cellular Dynamics International – a FUJIFILM Company, Madison, WI USA
- Scientific and Medical Affairs, CBR Systems, Inc., San Bruno, CA
Side by side comparisons of starting material for generating iPSCs have been conducted but often focus on adult tissues or tissue types from donors of varying ages. We previously demonstrated that iPSCs can be generated from umbilical cord blood processed with manual or automated platforms at a cord blood bank. Next we sought to compare the reprogramming capacity of multiple newborn tissue types from the same donor. iPSCs were generated from mesenchymal stem cells isolated from thawed umbilical cord tissue, previously cryopreserved as a composite material, using an integration-free reprogramming method. The resulting cord tissue derived iPSCs were characterized and the reprogramming efficiency and line quality compared to the cord blood derived iPSCs generated from the same donor. iPSCs were successfully generated from matched cord blood and tissue pairs from each of 5 donors. iPSC derivation from cryopreserved cord blood required less starting material than cord tissue. However, the resulting cord blood and cord tissue derived iPSC lines were equivalent in quality based on a panel of tests including pluripotency gene expression profiling, embryoid body formation, plasmid loss and karyotype analysis. This study confirms that cord tissue cryopreserved as a composite material in a cord blood bank is a suitable alternative source material to generate iPSCs. Having identified that donor specific cord blood and cord tissue iPSCs are comparable in quality is of interest in that it provides rationale for iPSC generation from a newborn tissue source processed and stored under relevant cord blood banking regulations while preserving the cord blood unit in its entirety for future clinical utility.