A versatile technological platform for the preparation of functionalized porous hydrogels detined to cell culture applications (INNOV Phase IA)

用于疾病研究、药物发现/功效和毒理学测定、** 干细胞/器官工程的大多数商业细胞培养材料是2D功能化表面。然而，已知它们促进细胞扁平化，导致与其体内环境相比显著的细胞行为差异。为了克服这个问题，用于细胞培养的3D多孔、功能化水凝胶材料已经开始出现在市场上。这样的材料将提供例如用于阿尔茨海默病或心脏病的药物筛选测定的更生理相关的工具。商业上可行的方法应该产生生物相容性材料，精确控制其3D微结构-孔径分布、体积分数、孔互连性以便于细胞接种并增强质量运输（营养物、生长因子、O2、废物等），并具有功能化的孔表面，以促进细胞粘附、存活和分化。该过程应可扩展，允许灭菌和长期储存。满足所有这些要求是一项挑战。据我们所知，市场上没有这种材料。这个I2 I 1A项目的主要目标是评估我们实验室开发的一种新技术的商业潜力，该技术用于设计和制备用于细胞培养应用的3D微结构化、功能化多孔藻酸盐凝胶，本文用间充质干细胞证明了该技术将分化为神经元（用于阿尔茨海默病研究）或心肌细胞（用于心脏组织损伤愈合的研究）。该方法使用商业加工聚合物设备来制备共连续的聚合物共混物，用于制备多孔聚合物，藻酸盐水凝胶被模制到该多孔聚合物中。提取模具产生多孔藻酸盐凝胶。我们的方法允许控制凝胶孔隙率：（i）40-60%的孔隙分数;（ii）10至500微米的平均孔径;（iii）完全互连的孔隙。具体目标是（1）优化多孔藻酸盐凝胶的制备;（2）优化凝胶功能化;（3）评估它们在细胞接种、粘附、存活和分化方面的性能;（4）开发灭菌/储存程序;（5）评估工艺和材料的商业潜力。

Most commercial cell culture materials for disease research, drug discovery/efficacy and toxicology assays,**stem cell/organ engineering, are 2D functionalized surfaces. However, they are known to promote cell flattening, resulting in significant cell behavior differences as compared to their in vivo environment. To overcome this problem, 3D porous, functionalized hydrogel materials for cell culture have started to appear on the market. Such materials will provide, for examples, more physiologically relevant tools for drug screening assays for Alzheimer disease, or cardiac pathologies. A commercially viable process should yield biocompatible materials, with precise control over their 3D microstructure - pore size distribution, volume fraction, pore interconnectivity to ease cell seeding and to enhance mass transport (nutrients, growth factors, O2, wastes, etc.), and with functionalized pore surface to promote cell adhesion, survival and differentiation. The process should be scalable, allow sterilization, and long-term storage. Meeting all of these requirements is a challenge. To the best of our knowledge, there is no such material on the market. The main objective of this I2I 1A project is to assess the commercial potential of a new technology developed in our laboratory for the design and preparation of 3D microstructured, functionalized porous alginate gels for cell culture applications, demonstrated herein with mesenchymal stem cells that will differentiate into neurons (for Alzheimer research) or cardiomyocytes (for research on healing of cardiac tissues injuries). The process uses commercial processing polymer equipment to prepare cocontinuous polymer blends, used to prepare porous polymers into which alginate hydrogels are molded. Extraction of the molds yields porous alginate gels. Our process allows control over gel porosity: (i) 40-60% pore fraction; (ii) 10 to 500 microns average pore size; (iii) fully interconnected pores. The specific objectives are (1) to optimize the preparation of porous alginate gels; (2) to optimize gel functionalization; (3) to evaluate their performance for cell seeding, adhesion, survival and differentiation; (4) to develop sterilization/storage procedures; (5) to assess the commercial potential of the process and materials.**********************