Project: Standardized 3D liver toxicity model: Combining unique hESC derived hepatocytes and scaffold technology

BACKGROUND_x000D_The biotechnological and pharmaceutical industries through-out Europe and ROW rely on animal-based systems for hepatotoxicity testing (liver toxicity), as an important part of almost every drug to be developed._x000D_Today, no commercially available standardized cellular models are adequate to predict human liver toxicity, and this segment is largely occupied by animal experimentation. However, animal models used are only about 50 % predictive, which is a huge problem in the drug discovery process and is causing pharma industry enormous capital loss due to late attrition rates. While human primary hepatocytes as a cellular model have proven to have drawbacks, human stem cell derived hepatocytes are expected to be commonly used in hepatotoxicity testing in the future, since they form three dimensional “organ-like” structures with improvement of important characteristics when cultured in three dimensional (3D) scaffolds compared to flat 2D cultures. In addition, human embryonic stem cell derived hepatocytes provide an unlimited source of homogenous and functional human hepatocytes._x000D_Suitable scaffolds for 3D cell culture are highly dependent on pore size, mechanical structure and spacing for liquid flow for proper nutrition medium and gas exchange. Various scaffolds have been developed independently by different companies, however the major challenge is to select the optimal scaffold for human embryonic stem cells (hESC), and to integrate these scaffolds in a cost-effective in-line production process into a micro-well plate format with small well sizes suitable for High Throughput Screening (>= 96 wells per plate). This will bring to market an commercially attractive standardized model for high throughput hepatotoxicity screening based on hESC, including related analysis methods for the 3D model._x000D__x000D_APPROACH_x000D_To develop highly innovative, unique and standardized model suitable for high throughput hepatotoxicity screening based on hESC, the complementary expertise of the three project Ps, two R&D performing SME's (Origo Biotech APS, Denmark and Cellartis AB, Sweden) and a technological service P, Bioneer in Denmark is required:_x000D__x000D_Origo Biotech holds the right to various scaffold and surface coating technologies as well as assembly method for integrating proprietary as well as commercially available scaffolds into multi-well plates in a in-line manufacturing process, including degradable and non-degradable scaffolds made by polystyrene, aluminum oxide and MPEG-PLGA._x000D_These techniques and technologies will be incorporated into the in-line production of the well plates containing suitable scaffold as a standardized high throughput container format. The advantages of these technologies are the ability to tailor-make scaffold specifications optimized for hESC derived hepatocytes, including control of material, pore size, scaffold thickness and wetability. The technologies further allows non-binding features of surrounding container surface preventing outgrowth of cells from scaffold into container, as well as cost efficient and standardized in-line scaffold immobilization methods into the well plates. _x000D__x000D_Cellartis is the only player in the world , who offer routinely produced and validated stem-cell derived hepatocytes as a commercial product. Cellartis have the capabilities to investigate whether scaffold materials are compatible with stem cell culturing as well as testing the optimal scaffold specifications during development including cell adhesion, cell death/survival, cell proliferation, differentiation and maturation. The company is the world’s largest single source of ethically derived hESC lines, and has expanded its operation into automated cell production and built the world's first production facility to make large volumes of hESC._x000D__x000D_Bioneers has expertise in stem cell cultivation and drug process development and validation. Therefore the consortium will have the capabilities to develop suitable analysis methods for cellular effects like changes in enzyme activity, bile canaliculi formation, cell polarization, expression of transporters in the membranes as well as to validation of cell models with existing drug. _x000D__x000D_EXPECTED RESULTS_x000D_The expected results include:_x000D_- The development of new 3D scaffold structures for 3D cell models in industrially accepted multi-well plate platforms._x000D_- The development and large scale manufacturing of a validated standardized liv_x000D__x000D_The direct beneficiaries will be the two participating SME's who expect a substantial company growth as a result of this project (Origo Biotech + 100% in employees), and Bioneer who will enrich their research competencies as well as service offerings to pharmaceutical industry. The indirect beneficiares will be the biotech and pharmaceutical companies who will benefit from the high through put screening services, i.e. faster and more realiable toxicity and screening analysis.

Acronym	HEPA-TOX (Reference Number: 5698)
Duration	01/08/2010 - 01/01/2013
Project Topic	Cellartis, Origo Biotech and Bioneer will develop the first high-throughput hepatocyte-based device for drug toxicity /hepatotoxicity testing combining human stem cell derived hepatocytes, scaffold technology as well as toxicity monitoring methods .
Project Results (after finalisation)	The CO results of the project is the implementation of analytical platforms for liver toxicity and metabolism as a Bioneer service.
Network	Eurostars
Call	Eurostars Cut-Off 4

Project partner

Number	Name	Role	Country
3	Bioneer A/S	Partner	Denmark
3	Cellartis AB	Coordinator	Sweden
3	Origo Biotech	Partner	Denmark