Reproducibility assessment of PeptiGel® products for stem cells culture and applications

干细胞是生物医学研究和应用的有力工具，包括复杂体外模型的产生、疾病建模、细胞治疗和药物发现。此外，干细胞可以分化形成不同类型的细胞，组织和器官，并可以生长以模拟疾病，如癌症和其他组织相关疾病。在干细胞环境中，分化受到细胞外基质成分和生长因子的影响，这些成分和生长因子提供了关键的指导信号。特别令人感兴趣的是它们在产生类器官（迷你器官）中的用途，其中干细胞簇聚集在一起以模拟组织和器官的天然微环境。使用干细胞的动机是它们改变我们对基础生物学的理解的非凡潜力，以及复杂的临床前模型的开发。它们还能够减少研究和开发中的动物使用。干细胞在三维（3D）中的生长和分化已在很大程度上通过使用市售动物源性产品（3D凝胶）得到证实。除了收获中涉及的伦理问题外，该产品还存在显著的批次间差异，这些差异来自个体间种属和供应商间差异。这种动物来源的3D凝胶还表现出热响应特性，在4 ℃下显示液体特性，但在室温下形成凝胶。这极大地影响了科学家如何使用它及其在高通量应用中的转化能力，因为这些参数与基本的高通量和机器人系统不兼容。因此，使用该产品的大多数实验都无法重现，数据解释也变得困难。为了充分了解干细胞的分化能力，并开发其减少研究中动物使用的能力，必须拥有一种非动物来源的3D凝胶，该凝胶具有一致的批量生产，易于处理的特性，以便在实验室或临床环境中更好地转化。我们的PeptiGel（r）产品是具有生物相关性的合成水凝胶，通过具有可调特性来模拟天然组织的天然细胞外基质，从而模拟组织的细胞环境。它们已被证明支持多种细胞类型的生长。然而，缺乏PeptiGels与干细胞群体相互作用的可靠数据。因此，本项目的目的是可重复地评估和验证PeptiGel（r）产品，以了解人类间充质干细胞培养的理想生长和分化环境条件。该项目的成功完成将推动PeptiGel（r）的市场渗透，进一步开发间充质干细胞工作的解决方案，并增加收入。

Stem cells are a powerful tool in biomedical research and applications, including generation of complex _in vitro_ models, disease modelling, cell therapy and drug discovery. Furthermore, stem cells can differentiate to form different types of cells, tissues and organs and can be grown to mimic diseases such as cancer and other tissue-related disorders. Within the stem cell environment, differentiation is influenced by extracellular matrix components and growth factors, which provide key instructive signals. Of particular interest are their use in generating organoids (mini organs), where clusters of stem cells come together to mimic the native microenvironment of tissues and organs.A motivation for the use of stems cells is their extraordinary potential of changing our understanding of basic biology, and the development of complex preclinical models. They also have the capacity to reduce animal usage in research and development. The growth and differentiation of stem cells in 3-dimensional (3D) have largely been demonstrated with the use of commercially available animal-derived product (3D gel). As well as ethical issues involved in its harvest, this product have significant batch-to-batch variability occurring from inter-individual species and inter-supplier variation. This animal-derived 3D gel also demonstrates thermo-responsive properties, displaying liquid properties at 4oC but forming a gel at room temperature. This significantly impacts how scientists work with it and its translational capacity in high-throughput applications, as these parameters are not compatible with essential high throughput and robotic systems. Most experiments using this product are therefore not reproducible and make data interpretation difficult.To fully understand the differentiation ability of stem cells and develop their capacity to reduce animal usage in research, it is imperative to have a non-animal derived 3D gel that shows consistent batch production, easy to handle properties for better translation in a laboratory or clinical setting. Our PeptiGel(r) products are biologically relevant, synthetic hydrogels that mimic a tissues' cellular environment through having tuneable properties to simulate the natural extracellular matrix of a native tissue. They have been shown to support the growth of a variety of cell types. However robust data of PeptiGels(r) interactions with stem cell populations is lacking. Therefore, the aim of this project is to reproducibly assess and validate PeptiGel(r) products to understand the ideal growth and differentiation environmental conditions for human mesenchymal stem cell culture. The successful completion of this project will drive forward PeptiGel(r) market penetration, further develop solutions for mesenchymal stem cell work and increase revenue.