Project: Optimal cell culture technology for applied stem cell development

The lack of effective, reproducible, animal-free hESC (human embryonic stem cells) and iPSC (induced pluripotent stem cells) culture methods, amenable to scale up presents a serious challenge to progression of new novel therapies into the clinical testing arena. It is furthermore critical for the development of robust reproducible in vitro cell based toxicology assay systems important to the development of new drugs. Thus, there is a growing need for cell culture technologies to support the specific needs of human ESC and iPSC cultures._x000D_Modification of surface materials with bioactive molecules for stimulated cell adhesion and growth has been investigated and reported with numerous findings in the fields of stem cell -and materials science. However as the needed signals for cells is a result of a variety of stimulations, optimal and robust coating technologies for incorporating such signals has not been identified. _x000D_The SURFStem Consortium Ps have identified a unique future market opportunity for creating an new combined cultivation and factor release technology for the functionalization of stem cell culture processes. _x000D_This combinatorial technology will bring several commercially attractive products on the market that; _x000D_1) will lead to optimized stem cell culture technologies that are able to support robust culture of various stem cell types._x000D_2) improve standardization of stem cell culturing processes - which is especially important in the growing field that relates to GMP regulated expansion of stem cells for clinical trials._x000D_3) reduce the cost of stem cell culture for the end user._x000D__x000D_APPROACH_x000D_To develop highly innovative, unique and standardized technology suitable for guided stem cell culture, the complementary expertise of the project Ps, two R&D performing SME's (Amphidex A/S, Denmark and Orla Protein Technologies Ltd, UK) and a technological service P, Bioneer in Denmark is required:_x000D__x000D_Amphidex holds the right to a novel technology for creating multilayer structures of cyclodextrins, that can be used for compound encapsulation. The novel Amphidex technology has been developed over the past five years and is now IP protected through several patents. Amphidex is capable of modifying these cyclodextrins for several purposes, including the combinnation with protein coating technology. _x000D__x000D_Orla Protein Technologies has developed a library of proteins that contain peptide sequences relevant to the culture of stem cells. The proteins are engineered to introduce doCOs or motifs from various bioactive proteins into a protein scaffold with self assembling properties that can be used to display the motifs in a correctly oriented, functional conformation on a variety of surfaces (gold, plastics, glass, silk, PLA/PLGA). The library contains a wide variety of short length peptide motifs from extracellular matrix proteins and also fusions of full length proteins and growth factors. _x000D__x000D_Bioneer has extensive expertise in applied stem cell culture, stem cell culture technology development and validation. Therefore the consortium will have the capabilities to conduct the necessary biological evaluation and testing of the developed technology for cell culture functionalization. Bioneer has state-of the art stem cell culture facilities and equipment for this part of the project and will generate data packages that is able to direct the product development. _x000D__x000D_EXPECTED RESULTS_x000D_The expected primary deliverable of this project will be a novel technology for functionalizing cell culture processes ready to be launched on the rapidly expanding stem cell market covering the two major areas within applied stem cells science.,_x000D__x000D_1. A robust and flexible technology for functionalizing already excisting cell culture surfaces leading to optimized stem cell culture _x000D_ - Cyclodextrin mediated encapsulation of chemical compounds, combined with protein modification of _x000D_ cell culture surfaces. _x000D_ _x000D_2. A robust and flexible technology for functionalizing cyclodextrins with proteins increasing the robustness and efficiency of the stem cell culture process. _x000D_ _x000D_The direct beneficiaries will be the two participating SME's who expect a substantial company growth as a result of this project (Amphidex + 100% in employees, Orla +20% increase in employees, +5% increase in turnover), and Bioneer who will enrich their research competencies as well as service offerings in relation to stem cell culture technology._x000D_The platform will also have direct beneficial effects on the life science industry and hospitals as the technology will ensure an efficient and robust expansion of stem cell populations.

Acronym	SURFStem (Reference Number: 8149)
Duration	01/09/2013 - 31/08/2015
Project Topic	Amphidex, Orla Protein Technologies and Bioneer will develop the first cell culture technology capable of delivering relevant biological signals to the cell culture in an efficient and low cost format.
Project Results (after finalisation)	1. Proteins (vitronectin) can bind to ACD on plast and glass surfaces, this was the initial PoC when the project started. Small molecules can bind to the ACD/protein-coat (proof of concept of the ACD Surface Coating™), in this case LDN and SB growth factors. This was also a step in the early compatibility studies during the PoC phase of the project. 2. The ACD coat is able to control the desired release concentrations of the small molecules LDN and SB, where a correlation between ACD amount is observed in the output concentration of released small molecules. 3. Furthermore it is observed and proven that the ACD coat binds the small molecules (LDN and SB) either by complexation or other surface electrostatic forces which produced a more slow release compared to standard plast surfaces - this property is relevant in the differentiation processes with iPSC's as a burst release of the small molecules is not desired. 4. A robust adherence and expansion of human induced pluripotent stem cells (iPSCs) was demonstrated and additionally it was demonstrated that iPSCs are capable of differentiating to neural stem cells on the combined coating of ACD and vitronectin, when neuronal inducers are added to the medium as in the conventional differentiation protocols
Network	Eurostars
Call	Eurostars Cut-Off 10

Project partner

Number	Name	Role	Country
3	Amphidex A/S	Coordinator	Denmark
3	Bioneer A/S	Partner	Denmark
3	Orla Protein Technologies Ltd.	Partner	United Kingdom