3D cell culture models are considered the next-generation in vitro models. Several efforts are being devoted to better mimic human physiology and predict drug efficacy. In case of the intestinal tissue models, the 3D architecture of the intestinal tissue and its soft mechanical properties have been identified as key factors to obtain models with improved predictive capacities. However, there is a lack of low-cost and ease-to-use intestinal tissue models that encompass both elements.
In the previous COMIET ERC project, we developed 3D cell culture scaffolds that accurately mimic the villus and crypt morphologies of the human intestine using soft materials. We used an innovative light-based approach to fabricate hydrogel scaffolds with the proper dimensions and mechanical properties of the tissue, in a reproducible manner. Once produced, the scaffolds were assembled in cell culture inserts (Transwells) to perform later assays. The technology was robust; however, the process of handling and transferring the scaffolds to standard cell-culture plates was manually preformed, increasing the total production time and the potential failures due to misalignments, sample contamination or even leakage.
In this ERC PoC GUT3D-PLATE we have moved a step forward improving and further developing the technology, to provide end-users with Ready-to-use cell culture plates for gut models, including individual scaffolds with the characteristic 3D intestinal physiology. These substrates are compatible with conventional assays and can be used without the need of specialized equipment. Moreover, they support the growth of conventional intestinal cell lines and primary-derived cells and can be used as platforms for drug screening, disease modeling and the study of microbiome interactions. Parallel production and process automatization have been our major goals. Indeed, we have designed an optimized manufacturing process to be able to print, individualize, and assemble with the minimum manipulation of the samples and the maximum yield. First, we have customized a 3D low-cost printing device to fabricate the hydrogel-based scaffolds by means of photopolymerization in a parallel manner. In particular, the system has been customized for printing of either 24 or 48 well insert plates at a time. To guarantee the permeation of nutrients and drugs, the bioprinted tissue-like hydrogels have been fabricated onto a porous membrane support, that later was assembled into a plastic cell insert to standardize the later culture. As initial step, single units and small arrays of up to 7 scaffolds were produced to prove the concept and optimize the different steps. So specific adaptors, printing tools and supports were manufactured accordingly. The individualization and attachment of the membranes to the Transwell inserts has been automatized customizing a low-cost pressing machine. Several supports and handling tools have been designed to perform both processes using the same device, ensuring proper alignment and reducing sample’s manipulation. Once ready, the sterility, packaging and storage conditions of the produced plates have been optimized thinking on their later delivery to costumers. Small series of ready-to-use GUT3D-plates were produced and distributed to different academic partners to test their suitability for different assays in house. Drug permeability, cell growth and host-pathogen interactions tests are examples of the assays performed, resulting in very positive feedback and thus, proving the potential value of our system and their potential interest as a commercial product. In this sense, we have also explored its commercial feasibility by protecting the IPR. A market analysis has been performed to identify the main product strengths, the potential costumers and key partners. In this sense, a questionnaire was distributed to get feedback and collect the main end-user requirements. Additionally, we several meetings with potential industrial partners were arranged to get close to industrial processes and obtain some specific information about processing and distribution. In parallel, a business plan was also developed, identifying the best roadmap to attract investors, taking into account the potential market competitors.
In summary, in the GUT3D-PLATE project we have developed a technology to produce testing platforms with complex 3D microstructures mimicking the gut, in a faster and robust manner. All the production steps have been carefully designed and adapted, allowing for a parallel production. By means of free-sample delivery to potential end-users, we have validated their suitability as ready-to-use platforms for intestinal 3D culture in biomedical research, also proving their viability for the pharma industry and CROs, due to the increasing demand of tools that better predict drug treatment efficacy and toxicity at early stages of the drug development processes. Our GUT3D-PLATE testing platforms significantly reduce the need for animal models and the overall cost, generating a major societal benefit.
