Using iPS cell derived sensory neurons in predicting drug-induced peripheral neurotoxicity
Peripheral neurotoxicity is a major side effect of chemotherapy agents but also across a wide range of other drugs. Safety toxicity is a leading cause of drug failures both pre-clinical and during clinical development. Effective testing of drugs for neurotoxicity has been hampered by a lack of relevant in vitro models.
Dr Sarah Harper, Principal Scientist at Axol Bioscience, analysed at the "Good Cell Culture Practices" seminar in Vienna why human iPSC-derived Sensory Neurons can be a reliable in vitro model that could be used in predicting drug-induced peripheral neurotoxicity.
Pharmacological evaluation in human iPSC-derived cortical and sensory neurons using high-throughput MEA system
Human iPSC-derived neurons are expected as a new toxicological evaluation assay to replace animal experiments in preclinical studies, improving the risk of accuracy.
The most sensitive microelectrode array system for in vitro extracellular electrophysiology
MEAs have been adapted to drug discovery assays including cell-to-cell interaction, learning and memory, and network properties of neurons. High sensitivity consists key factor of their performance; it results in better signal to noise ratio leading to more reliable detection of action porentials. Michael Trujillo, Product Manager at Alpha Med Scientific, explains how low noise in engineered into MED64 systems.
Frontotemporal dementia and Parkinson's Disease cellular models: In vitro characterization of pathological phenotypes in gene-edited iPSC-derived neurons with MAPT and LRRK2 mutations
Mutations in MAPT and LRRK2 are reported to play a causative role in familial forms of frontotemporal dementia and Parkinson’s disease respectively. Human cell-based models in which to investigate the effect of these mutations on cellular functions such as microtubule assembly, oxidative stress, mitochondrial- and synaptic-functions, all of which could contribute to the onset of neurodegenerative conditions, are necessary to understand the disease mechanisms. Genome-edited human iPSC-derived neural stem cells (NSCs) offer a virtually unlimited source of physiologically relevant isogenic cell lines for use in disease modeling and drug discovery. Combining iPSC genome editing using CRISPR-Cas9 and directed differentiation, we have generated patient-relevant NSC disease models (axolGEMs) carrying heterozygous and homozygous combinations of missense mutations R406W, P301L and V337M in MAPT and G2019S in LRRK2. Genotype, karyotype, SNP frequency and copy number were assessed in genome-edited iPSC lines. Differentiated cells formed polarized neural tube-like rosettes in a monolayer culture. Immunocytochemistry confirmed the expression of typical NSC (PAX6, FOXG1, nestin, Ki67 and NCAD) and cerebral cortical neuronal markers (TUJ1, TBR1, MAP2 and CTIP). These cells enable a direct comparison of the variant effect on cellular phenotype between isogenic lines and offer a stable platform for drug screening and validation.
Abnormal phosphorylation of amyloid precursor protein tyrosine residues alters the APP trafficking in neurons from Alzheimer's Disease affected patients
Alzheimer’s Disease (AD) is the most common cause of dementia and is likely caused by defective amyloid precursor protein (APP) trafficking and processing in neurons leading to amyloid plaques containing the amyloid-β (Aβ) APP peptide byproduct. Understanding how APP is targeted and trafficked to selected destinations inside neurons and identifying the mechanisms responsible for the generation of Aβ are thus the key for the development of new therapies. We previously developed a mouse model showing that the Tyrosine (Tyr)682, in the C-terminus of APP, is essential for its binding to the coating protein clathrin and to the clathrin adaptor protein AP2 and for APP trafficking and sorting in neurons. In the present study, we investigated whether the Tyr682 residue of APP influences APP trafficking and sorting in neurons from differentiated neural stem cells (NSCs) of patients with AD carrying mutations on the PS1 gene (L286V; A246E; M146L), and on cortical tissues and fibroblasts from Göttingen minipigs engineered to develop AD (M146I mutant minipigs). Our results show that Tyrosine phosphorylation controls the APP binding to clathrin and AP2 in neurons carrying PS1 mutation and influences APP trafficking and sorting. Overall, our results provide a basis for the development of potential new therapies for AD.
Modeling FTDP-17 linked tauopathies and Alzheimer's disease with human iPSC
Dr An Verheyen, Janssen R&D Discovery Services presented her work at ISSCR Basel, 2017. Dr Verheyen used human iPSC-derived neural cells generated using our custom differentiation service to model tauopathies linked to frontotemporal dementia with parkinsonism-17 and Alzheimer's disease.
Electrophysiological maturation and pharmacological responses of human induced pluripotent stem cell-derived cortical neuronal networks in long-term culture
Ikuro Suzuki, Associate Professor at Tohoku Institute of Technology, Japan who recently published his work with Axol Human iPSC-Derived Cerebral Cortical Neurons in Nature’s Scientific Reports (Odawara et al., 2016), presented our Innovation Showcase in collaboration with Alpha MED Scientific.
Prof Suzuki discussed the following results from his research:
- Morphology in long-term culture
- Development of spontaneous burst firings
- Pharmacological properties
- Induction of epileptiform activity & effects of anti-epilepsy drugs
- Induction of long-term potentiation and depression
Modeling Alzheimer's Disease
Dr Alfredo Cabrera, Janssen Pharmaceutica, presented a robust, scalable and disease relevant model of tau aggregation using induced pluripotent stem cell (iPSC)-derived cortical neurons that can be applied to drug discovery programs in neurodegeneration. The resulting assay is highly reproducible across users and works in different commercially available iPSC-lines, providing a reliable tool for better understanding TAU pathophysiology and the identification of novel treatments against Alzheimer’s disease.
Modeling Alzheimer's disease using stem cells
Currently, the majority of studies on Alzheimer's disease have used transgenic animal models or imaging studies of the human brain. It is difficult to validate these findings using human tissue. Whilst animal models have been central to our understanding of human physiology, human stem cell-based models may help us to further our understanding of human physiology and tackle devastating diseases such as Alzheimer's disease.
Induced pluripotent stem cell-derived endothelial colony forming cells offer a robust and physiologically relevant research tool
Human iPSC-Derived Endothelial Colony Forming Cells (ECFCs) (Axol Bioscience) are highly expandable and show comparable expression and functionality to primary cells, providing a robust and physiologically relevant tool for use in numerous applications. ECFCs are rare circulating endothelial cells that display a hierarchy of clonal proliferative potential and possess in vivo vessel-forming ability upon implantation. In numerous animal models of disease, human ECFCs have demonstrated the capacity to promote revascularization and reperfusion to injured vascular beds via direct integration and/or through paracrine effects.
Human umbilical cord blood is enriched in circulating ECFCs compared to adult peripheral blood. We have recently reported that ECFCs displaying properties similar to cord blood can be obtained from human ESCs and iPSCs. Using defined serum-free culture medium and sequential addition of specific growth factors, we’ve identified ECFC precursors within 12 days of iPSC differentiation. These iPSC-derived ECFCs display clonogenic proliferative potential and in vivo vessel forming ability similar to cord blood ECFCs and promote vascular repair and regeneration in multiple animal models of human disease.
Human iPSC-derived cardiomyocytes: A comparison with primary cells and applications in standard and 3D culture models
The University Hospital Bern studies the mechanisms of cancer therapy-associated cardiotoxicity, and wants to study this in cardiomyocytes, thus they always endeavor to use the most relevant in vitro culture systems. Therefore, they have recently started to develop a 3D-culture model using hiPSC-derived cardiomyocytes and have tested this system in comparison with mature primary cells.
Stem Cell-Derived Human Renal Cells for Compound Screening and Nephrotoxicity Prediction
The development of the first pre-validated screening platforms for accurate prediction of nephrotoxicity in humans, and the potential applications of these cells within drug discovery and toxicology with progression to future kidney-on-chip technologies for dose response prediction.
The use of human iPSC-derived renal proximal tubular cells in predicting nephrotoxicity
The innovation of human iPS cells has revolutionized the field of drug discovery and development, offering researchers access to a consistent and almost infinite supply of stem cells from a single donor source. Human iPSC-derived cells can offer a more physiologically relevant model to be used for screening candidate drugs. Human iPSC-derived renal proximal tubular cells are an example of a new cellular development that would be ideal for use in drug discovery to identify and predict drug induced toxicity.
Stem cells: opportunities, hurdles and promises
Dr Matthias Stadtfeld, Assistant Professor at the New York University School of Medicine and Dr Kejin Hu, Assistant Professor at University of Alabama at Birmingham, discussed the potential of stem cells for medical research and drug discovery.
The webinar organised by The Scientist and supported by Axol Bioscience analysed further the molecular mechanism of induced pluripotency as well as the roadblocks to iPSC repogramming and erasing transcriptional memory in cellular reprogramming.
Relevance of iPS cells in cell based screening and drug development
Drug development requires highly validated, reproducible, consistent and predictive models in order to quickly validate key targets and assess toxicity in pre-clinical assays.
High quality human iPSC cells can help to that direction and Nick Clare analyses further in his presentation at ELRIG Drug Discovery 2017 how that can be achieved.
An overview of Axol Bioscience
Axol Bioscience specialize in the supply of human cell culture systems and custom services for disease modeling and drug discovery. Our expertise includes induced pluripotent stem cell (iPSC) generation, CRISPR-Cas9 gene editing, iPSC differentiation and custom cell and tissue sourcing.
We offer a variety of functionally validated human primary cells and human iPSC-derived cells from both healthy and patient donors alongside culture media and reagents that are tailored to support the optimal growth and maturation of our cells. This enables you to carry out your project in a physiologically relevant system that is not only reliable and easy to use but produces consistent results for robust replication studies.
Our team is passionate about great science, delivering superb customer service and support and innovating future products to help customers advance their research faster.
At Axol, innovation, quality and customer service are key.
The use of iPSC-derived cells and primary cells as in vitro models for toxicity screening
Toxicologists have access to a range of iPSC-derived cell types, including cardiomyocytes, hepatocytes and renal cells, used in toxicity screening. We discuss how these models are accurate and representative cell models, and how they can phase out inconsistencies and reduce the use of in vivo models.
Rising to the challenges of human iPSC-derived cells for tox & drug screening
We presented 'in the field' data on our portfolio of human iPSC and primary cells demonstrating their proven ease of use, reliability and consistency as meaningful drug discovery tools. Here, we overcome the challenges of cell line variability and address the needs for cell scale-up in assay campaigns.