MipTec 2016

 MipTec 2016 took place from 20 - 22 September during Basel Life Science Week. View our posters and presentation below.


Posters

Serum-Free Human iPSC-Derived Cardiomyocytes for in vitro Testing

Using defined factors OCT3/4, KLF4, SOX2 and c-MYC, adult cells from healthy and patient donors can be reprogrammed to generate induced pluripotent stem cells (iPSCs). Subsequently, these can be differentiated into a variety of cell types including cardiomyocytes. Human iPSC-derived cardiomyocytes (iPSC-CMs) can be cultured in vitro under serum-free conditions and as such, offer a platform investigate the effect of growth factors, cytokines and drugs on the development and functionality of human cardiomyocytes in vitro. Following differentiation, spontaneously beating iPS-CM’s were evaluated in 2D and 3D culture. Expression profiling by immunohistochemistry confirms the expression of cardiomyocyte selective markers including α-actinin, myosin heavy chain, atrial and ventricular myosin light chains, troponin-T and -I, β-catenin, vimentin, L-type calcium channels, connexin-40 and -43, telethonin and ankyrin repeat domain-1 (ANKRD1). Immunohistochemistry findings were validated by Western Blot for α-actinin and cardiac troponin-T expression. Additional analyses conducted, include bi-nucleate cell counts, cell form factor measurements and a comparison of plating efficiencies across a variety of substrates. The electrical activity of the iPSC-CMs was confirmed using a multi-electrode array (MEA), and the calcium dye Fluo4. One application of these cells is drug toxicity testing. To show proof of principle that this can be undertaken in a contactless manner using only genetically encoded tools, which offers several advantages compared to low throughput contact based methods with chemical dyes, we developed a simultaneous optical control/calcium imaging approach to replace the need for electrode stimulation and dyes. We are able to control the beat frequency of iPSC- CMs across the physiological range (0.3Hz – 2Hz) and can observe the anticipated effects of compounds such as Dofetilide, a known hERG inhibitor. Here, we have identified a range of characteristics in these human iPSC-CMs that confirms their ability to function as a highly-pure population of single beating human cardiomyocytes in vitro and presented evidence of a technically simple and scalable platform for cardiotoxicity screening assays.

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Pharmacological Responses in Cultured Human iPSC Derived Cortical Neurons Using Multi-Electrode Array

The functional network of human induced pluripotent stem cell (hiPSC)-derived neurons is a potentially powerful in vitro model for evaluating disease mechanisms and drug responses. However, the culture time required for the full functional maturation of individual neurons and networks is uncertain. We investigated the development of spontaneous electrophysiological activity and pharmacological responses for over 1 year in culture using multi-electrode arrays (MEAs). The complete maturation of spontaneous firing, evoked responses, and modulation of activity by glutamatergic and GABAergic receptor antagonists/agonists required 20–30 weeks. At this stage, neural networks also demonstrated epileptiform synchronized burst firing (SBF) in response to pro-convulsants and SBF suppression using clinical anti-epilepsy drugs. Our results reveal the feasibility of long-term MEA measurements from hiPSC-derived neuronal networks in vitro for mechanistic analyses and drug screening. However, developmental changes in electrophysiological and pharmacological properties indicate the necessity for the international standardization of culture and evaluation procedures.

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Implementation of High Throughput Screening on iPS Cells

Induced pluripotent stem cells (iPSCs) belong to a growing list of stem cell populations that hold great potential for use in cell-based assay development and screening. iPSC-derived cells like cardiomyocytes and neuronal cells represent a reliable source of cellular material with robust assay performance and scalability which is essential to any HTS campaign and subsequent follow-up. Here we demonstrate the impact of iPSC-derived neurons in modern drug discovery. We present examples of assay automation and miniaturization of cellular assays based on high content imaging using the Operetta and Ca-flux using the FLIPR instruments. Our examples show the opportunities and benefits that iPSC-derived cells may provide in the search for new chemical starting points for drug discovery in the near future.

20 Qualified Hit List reports to the target owners. The over 40 HTS campaigns performed to date cover a wide range of target

classes including more demanding cellular targets like ion channels and GPCR’s. Cellular HTS assays have been successfully

miniaturized to 384- and 1536-well format in order to perform them in a cost-efficient manner. Induced pluripotent stem cells

(iPSCs) belong to a growing list of stem cell populations that hold great potential for use in cell-based assay development and

screening. iPSC-derived cells like cardiomyocytes and neuronal cells represent a reliable source of cellular material with robust

assay performance and scalability which is essential to any HTS campaign and subsequent follow-up. Here we demonstrate

the impact of iPSC-derived neurons in modern drug discovery. We present examples of assay automation and miniaturization

of cellular assays based on high content imaging using the Operetta and Ca-flux using the FLIPR instruments. Our examples

show the opportunities and benefits that iPSC-derived cells may provide in the search for new chemical starting points for drug

discovery in the near future.

 

Read it now


Presentations

Human iPSC-Derived Cardiomyocytes - A Comparison With Primary Cells and Applications in Standard and 3D Culture Models

Dr Christian Zuppinger (Bern University Hospital, Switzerland) presented data on our Human iPSC-Derived Ventricular Cardiomyocytes including cardiac-selective marker expression, safety pharmacology testing results and 3D culture of scaffold-free micro-tissues aggregated in the hanging drop with/without other cardiovascular cell types.

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