Unfortunately, side effects of active substances are often part of everyday life in pharmaceutical studies. As a result, serious complications can arise in daily clinical routines. Toxic effects, especially in the cardiac and neuronal area, are the main reasons why promising compounds must be withdrawn in late preclinical studies or even after market approval. State of the art in drug research are still in vitro cell culture models based on animal cells or in vivo animal experiments. Both systems can usually only be transferred inadequately to the human system, with the result that side effects are only discovered at a late stage and result in both economic and health damage. Therefore, there is an urgent need for significant test systems that allow a reliable prediction of possible drug-induced toxicities in humans.

Use of specialized cell types from patient-specific stem cells

The discovery of human induced pluripotent stem cells (hiPS cells) with their unique potential for unlimited self-replication and their ability to differentiate into almost all cell types of the human body led to major technical advances in the field of human cell-based toxicity studies. This cell system can, for example, be established from adult skin biopsies and then differentiated into different cell types, e. g. heart muscle or nerve cells. In this way, entire patient groups can be easily but specifically mapped in the laboratory and the cell types relevant for toxicity studies can be generated. At present, however, the processes involved in working with these cell systems are still characterized by the need for a great deal of practical expert knowledge and a great amount of time. Furthermore, these "assays" have to be produced on demand, can only be stored within a very limited time frame of a few days and are not transportable in their physiological state. Conventional cryobiological processes, i.  e. the controlled cooling of biological samples below freezing point, are already established for single cells in suspension. However, they cannot be used for the assay-relevant specific cell types cardiomyocytes and neurons, which grow on surfaces and only thus develop their functionality.

Fraunhofer IBMT expertise: Application-oriented cryopreservation through vitrification

The Fraunhofer Institute for Biomedical Engineering IBMT has decades of experience in cryobiotechnology and biobanking. Within the joint project "R2U-Tox-Assay" the Fraunhofer IBMT has taken on the task of generating functional cardiomyocytes and neurons from hiPS cells in scalable processes and freezing them in standardized laboratory plate formats for high-throughput screenings. The functionality of these cells is guaranteed by their contact with specially coated surfaces, which must be maintained during the cooling process. Ultra-high cooling rates and a special cocktail of cryoprotective agents ensure cryogenic temperatures below 140 °C that is reached without the formation of potentially damaging ice crystals within the sample. In this temperature range the samples can be stored and transported for an unlimited period of time. Immediately after thawing, they can be used in standardized assays without the time-consuming recovery phases associated with conventional cryopreservation.

In "R2U-Tox-Assay", the three participating partners combine their respective expertises. The Fraunhofer IBMT is coordinating the project and is responsible in particular for producing significant human cell systems and freezing them in application-oriented formats. Janssen Pharmaceutica N.V., part of the Janssen Pharmaceutical Companies of Johnson & Johnson, validates the developed test systems in the pharmaceutical environment in already established compound screenings. The Institute for Bioengineering of Catalonia establishes genetically engineered so-called reporter cell lines, which allow a faster analysis of potentially toxic effects within the cell system. 

Project Funding: EIT Health INNOVATION Project

Funding period: 01/2020-12/2023

Project coordination:
Fraunhofer Institute for Biomedical Engineering IBMT, Sulzbach, Germany

Project partners:
Janssen Pharmaceutica N.V., Beerse, Belgium, part of the Janssen Pharmaceutical Companies of Johnson & Johnson (Project leader: Dr. Juan Pita Almenar and Dr. Hua Rong Lu)
Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain (Project leader: Dr. Nuria Montserrat)

Project manager at the Fraunhofer IBMT:
Dr. Julia Neubauer
Head of Department Cryo & Stem Cell Technologies
Head of Project Center Stem Cell Process Technology at the Fraunhofer ISC, Würzburg
Phone: +49 931 4100 360
E-mail: julia.neubauer@ibmt.fraunhofer.de

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EIT Health is one of the currently eight funded Knowledge and Innovation Communities (KICs) of the European Institute of Innovation and Technology (EIT). EIT Health delivers solutions for high-performing health systems, healthier citizens and a sustainable health economy in Europe. It does this by connecting the right people with the major health challenges of our time across European borders. EIT Health leverages the expertise of more than 150 leading players in European healthcare – pharmaceuticals, medical technology, diagnostics, consumer goods, universities, research institutions, healthcare providers, and payers, policymakers and administrators - to make innovation a reality and commercialise patient-oriented solutions.