27. - 31.03.2022 | SOT: 61st Annual Meeting and ToxExpo
San Diego Convention Center
111 West Harbor Drive
San Diego, CA 92101
Go to the Conference website here.
Session Date and Time: Monday March 28th, 2022 - 10:45 AM - 12:30 AM PST
Abstract Title: Tackling Human iPSC-Derived Cardiomyocyte Maturation In Vitro for Predictive Nonclinical Cardio-Toxicological Assessment (innoVitro collaboration)
Presenting Author: Wyatt Hall (Director of Sales - North America, Nanion Technologies)
Session Title: Stem Cell Biology and Toxicology
In pre-clinical drug development, human induced pluripotent stem cell-derived cardiomyocytes (hiPSC CMs) serve as animal-free approach to assess preclinical cardiac risk without ethical concern but high human relevance. However, their premature phenotype in standard cell culture assays leads to limited use in preclinical drug development. The bio compliant 96-well FLEXcyte technology, containing flexible membranes instead of stiff plastic surfaces, offers a physiological environment by mimicking a native-like environment for the cells. To fully analyze the effect of the FLEXcyte technology on hiPSC-CMs maturation, expression levels of relevant cardiac genes and phenotypic characterization via actin cytoskeleton immunostainings were performed as well as functional characterization with cardio-safe and cardio-toxic gold standard compounds. In addition, long-term culture of hiPSC-CMs on FLEXcyte plates were conducted and analyzed regarding changes in amplitude and beat rate over time. The results show a clear pro-maturation effect on functional properties of hiPSC-CMs when cultured on FLEXcyte 96 plates. Compounds considered as clinically cardio-safe showed negative inotropic effects only at high (micromolar) doses, while compounds with known cardiotoxic profiles showed both time and dose dependent inotropic effects. Changes in expression levels of cardio-specific genes as well as pronounced filamentous actin underlines the pro maturation effect on genetic and phenotypic level. Long-term culture over 44 days showed a significant increase in amplitude along with a decrease in beat rate, both known parameters for cardiac maturation. These results indicate that the FLEXcyte technology facilitates hiPSC-CM maturation not only on a functional but also a molecular and phenotypic level, hence providing a comprehensive solution for the assessment of human-based preclinical cardiac risk.
Session Date and Time: Wednesday March 30th, 2022 - 10:45 AM - 12:30 AM PST
Abstract Title: Comprehensive Impedance-Based Hepatotoxicity Assay for Metabolically Active IPSC-Derived Hepatocytes (FUJIFILM Cellular Dynamics, Inc. collaboration)
Presenting Author: Ronald Knox, PhD (Account Executive, Nanion Technologies)
Session Title: Liver II: In Vitro
Drug-induced hepatic toxicity is one of the main reasons for regulatory actions and market withdrawals of drugs in the last 50 years, while Drug-Induced Liver Injury (DILI) still remains the major cause of acute liver failure. Current existing in vitro models employed to predict DILI mostly focus on hepatocytes. iCell® Hepatocytes 2.0 (FUJIFILM Cellular Dynamics) are human iPSC-derived cells with a wide variety of basic and functional characteristics which make them amendable to a number of applications such as compound mediated ADME-T and DILI toxicity. iCell Hepatocytes 2.0 display characteristic hepatocyte morphology, including polygonal shape, polynucleation and formation of bile canalicular channels. These cells have been extensively characterized for expression of the liver cell markers including, albumin, A1AT, and HNF4a, as well as basic and induced P450 functions, as observed in primary human hepatocytes. The impedance signal recorded using planar gold-film electrodes reveals alterations in confluency, cell contact (morphological shape) and conductivity of adherent cells. thereby provides a measure of toxicity. This measurement gives valuable insights in various cell phenotypes, even over prolonged periods of time (acute and chronic exposures). Here, iCell Hepatocytes 2.0 were recorded on the CardioExcyte 96 (Nanion Technologies) and changes in impedance, therefore confluency, were used to measure of DILI. Acute and chronic (dose-dependent) liver toxic effects have been tested for 5 relevant compounds: diclofenac, APAP, Troglitazone, Chlorpromazine and Aflatoxin B1. To obtain 2D monolayers in 96-well plates, cells were seeded with density of 300k cells/cm2 , and recordings were performed during the time period of 21 days. Data showed comparable results to other cell viability and tox assays, such as CellTiter-Glo, rendering impedance as a reliable but non-invasive tool. We were able to show dose and time dependant effects for all 5 compounds on iCell Hepatocytes 2.0. Importantly, the acute and chronic effects of Aflatoxin B1 were observed in standard 2D conditions, indicating that the hepatocytes were metabolically active and functional when using this impedance-based, non-invasive assay.