ISSCR 2021 Virtual
The ISSCR Annual Meeting gathers nearly 4,000 of the world's leading scientists, clinicians, business leaders, ethicists, and educators from more than 60 countries. Together, we represent the full spectrum of stem cell science and regenerative medicine. Discovery, new business, careers, advocacy, and relationships start here.
FCDI Talk: Common Non-Scientific Challenges in The Generation, Use and Sharing Of iPSC Lines
Speaker: Dr. Eugenia Jones, Vice President, Head Life Sciences Strategic Office
Date: June 21
Time: 13:00-16:00 EDT
Abstract: The European Bank for iPSCs (EBiSC) is a centralised repository, currently in a second project phase including both non-profit and commercial iPSC researchers (EBiSC2), working to make iPSC tools available and developing protocols which improve and simplify their use. This focus session will share how EBiSC2 partners are adapting and consolidating iPSC expansion, differentiation and cryopreservation approaches to help ease transition into high volume applications whilst also ensuring accessibility for non-expert users. We will discuss how the inclusion of iPSC tool lines in these protocol developments enables rapid generation of functionally mature derived cell types and how the associated iPSC datasets can be broadly shared in an ethically compliant manner. Lastly, common stumbling blocks will be discussed to raise awareness across the community.
Date: June 21 Time: 13:00-16:00 EDT
Other Session Speakers: Julia Neubauer, PhD, Fraunhofer-IBMT, Germany; Mikkel Rasmussen, PhD, Bioneer, Denmark; Emilie Lemesre, PhD, Servier, France; Alfredo Cabrera-Socorro, PhD, Janssen Pharmaceutica NV, Belgium; Benjamin Schmid, PhD, Bioneer, Denmark Andreas Kurtz, PhD, Fraunhofer-IBMT, Germany
FCDI Poster Presentations
Investigating the Role of the AD Protective CD33 Variant in the Context of APOE Genotype Using Human iPSC-Derived Microglia
Poster Number: 753
Poster Session 7, 6/25/2021, 12:00- 13:00 EDT
Authors: Sarah Burton, Christie Munn, Madelyn Goedland, Mai Xiong, Simon Hilcove, Wayne Poon and Deepika Rajesh
Abstract: Recent Genome Wide Association Studies (GWAS) have identified a number of single nucleotide polymorphisms (SNPs) within genes expressed in human microglia that modify late-onset Alzheimer’s Disease (LOAD) risk. Inheritance of the e4 allele of Apolipoprotein E (APOE4) or the R47H mutation in triggering receptor expressed on myeloid cells 2 (TREM2) increase risk, while the rs12459419 T variant in CD33, a gene that encodes a member of the Sialic acid-binding immunoglobulin-like lectin (Siglecs), is protective. Given the genetic link between APOE4 and CD33 on LOAD and a strong correlation between the copy number of the protective CD33 rs12459419T allele with the dose-dependent decrease in AD risk, human induced pluripotent stem cells (iPSC)-derived microglia carrying these different SNPs were generated to understand the synergistic contribution of these variants in the development of AD. Episomally reprogrammed iPSCs from (i) healthy donors expressing APOE3/3 genotype and (ii) AD donors expressing an APOE4/4 genotype with the CD33 protective rs12459419T allele or the non-protective rs12459419C allele were expanded and successfully differentiated into microglia. Cryopreserved microglia from all donor samples expressed microglia-specific cell markers (CD45, TREM2, CD33, P2RY12, TMEME119, CX3CR1, IBA-1). Functional assessment of end stage cryopreserved microglia identified altered phagocytosis kinetics and differences in soluble TREM2 levels between donors harboring either the protective rs12459419T or the rs12459419C allele. Additional functional characterization was performed by stimulating microglia with either LPS (M1) or IL-4 + dBu-cAMP (M2) followed by quantification of the analytes. Microglia derived from donors harboring the protective rs12459419T allele released higher levels of immunomodulatory M2 analytes including IL-10, IL-13, IL-12, IL-27, CCL8, CCL13 and CCL6 compared to microglia harboring the non-protective rs12459419C allele in donors harboring APOE 3/3 versus APOE4/4. These findings unveil the mechanism of the cellular responses elicited by the protective rs12459419T allele in the context of APOE genotype. This panel of iPSC-derived microglia can be used to increase the understanding of the interplay of multiple genetic variants in modifying AD risk and provide valuable therapeutic targets for AD treatment.
Scalable Expansion Culture of Human Induced Pluripotent Stem Cells Using Microcarriers
Poster Number: 753
Poster Session 7, 6/25/2021, 12:00- 13:00 EDT
Authors: Tomoaki Kurakazu1, Sho Sato1, Ryo Hibino2, Maki Mifune2, Takahiro Oba2, Hideaki Kagawa1, Nathaniel Beardsley1, Kiranmayee Bakshy1, Sarah Dickerson1, Chad Koonce1, Peter Fuhrken1
1. FUJIFILM Cellular Dynamics, Inc
2. FUJIFILM Corporation, Bio Science & Engineering Laboratory
Abstract: Human induced pluripotent stem cells (hiPSCs) have unlimited potential for cell therapy products to address unmet medical needs and disease modeling tools to unlock new possibilities for drug discovery. A scalable and reproducible cell expansion process is important for the translation from research to commercial manufacturing. Moreover, for clinical application, it is advantageous to use a single-use, closed and controlled environment. At FUJIFILM Cellular Dynamics, Inc. (FCDI) , a leading developer and manufacturer of hiPSC derived cells used in drug discovery, toxicity testing, stem cell banking, and cell therapy development, we have experience in process optimization with various culture formats. Here, we present microcarrier-based hiPSC three-dimensional (3D) culture up to 10L scale. Under optimized conditions with vertical wheel mixing, we achieved up to 10.4 billion cell yield after 9 days while maintaining comparable cell quality including pluripotency marker expression and viability. In addition, closed cell washing and concentration by counterflow centrifugation was demonstrated with high viability and a recovery ratio over 85%. Post-processed iPSCs were successfully tested in multiple downstream applications including plating into 2D, making 3D aggregates for differentiation, and cryobanking. Together, these established iPSC scale-up and processing methods allow for the routine manufacture of > 10 billion iPSCs per batch.
Other Poster Presentations
Microfluidic High-Throughput Screening Platform to Screen Pre-Clinical Stage Compound Effects on Neurite Outgrowth of Human Motor Neurons Post Injury
Poster Number: 222
Poster Session 2, 6/23/2021, 00:00- 01:00 EDT
Authors: Dr Jessica Rontard, The Neuro Engineering Technologies Research Institute (NETRI)
Abstract: Traumatic spinal cord injury (SCI) affects millions of people of all ages around the world and can potentially lead to irreversible cognitive and motor damages as no relevant therapeutics are available. There is a significant demand for physiologically relevant models of SCI. The rise of Organ-on-Chip technologies combined with human cells opens new route to limit the number of animal experiments and create physiologically relevant disease model. Organ on chip technology offers the possibility to perform an an accurate injury on the neurites, without affecting cell viability and to apply the pre-clinical stage compounds on cell soma only. We will present the design and fabrication of a compartmentalized microfluidic device with three compartments capable of both inducing a localized axotomy and a fluidically isolated cell chamber. The device allows quantification of axonal regeneration post-injury using an innovant and unique triangular design. Human-induced pluripotent stem cell-derived motor neurons are seeded and maintained up to 3 weeks in vitro. We performed a chemically-induced axonal injury on isolated axons in the middle compartment only, opening the way to test neuropharmacological compounds, solely in the soma chamber. We show that this model can be successfully used for quantitative analysis of neurite outgrowth dynamics. We will finally present how this model could be used to record the functional activity using multielectrode arrays (MEA) coupled to the microfluidic device. The access to functional data during the injury and the recovery process provides relevant insight on the mode of action of pharmacological compounds. Our data suggest that this model can be used for high-throughput drug-induced axonal regeneration screening for pre-clinical stages pharmaceutical compounds.
Event Date: 2021-06-21 To 2021-06-26
Organiser: The International Society for Stem Cell Research