A Novel Defined Xenobiotic Free Substrate that Mimics Natural Extracellular Matrix

描述(申请人提供)：Primorigen Biosciences SBIR建议摘要Primorigen Biosciences将开发一种明确的、无异物的合成细胞培养基质，该基质完全支持人类诱导多能干细胞(HiPSCs)沿所有三个胚层的维护和定向分化，并显著降低干细胞培养过程的成本。目前最常用的基质要么产生严重的可靠性和安全性问题，要么仅对某些细胞系有效，因为它们的细胞黏附机制有限。为了解决这些缺陷，Primorigen将开发一种新的合成细胞培养基质，通过提供各种合成配体来概括天然细胞外基质的关键属性，这些配体提供多种细胞黏附机制，并促进内源性因子的隔离，从而减少使用外源生长因子的需要和成本。第一阶段研究将通过(A)创建合成细胞黏附和内源性因子隔离配体的库，(B)优化配体混合物在聚苯乙烯表面的涂层条件，以及(C)验证新的合成细胞培养基质，以维持hPSC并定向分化为各种谱系，从而证明其可行性。第二阶段的目标将建立在这项工作的基础上，通过扩大合成配体混合物的方法来验证它们在大规模生产和分化HiPSCs中的使用。商业产品将包括(I)广泛适用的合成Matrigel替代品，包括针对干细胞维护和分化而优化的合成模块配体混合物，(Ii)预包覆有合成模块配体混合物的聚苯乙烯组织培养平台(如培养皿、多孔板和微载体)，以及(Iii)合成模块配体资料库，从中可以根据客户的独特需求定制定制的混合物。

DESCRIPTION (provided by applicant): Primorigen Biosciences SBIR Proposal Abstract Primorigen Biosciences will develop a defined, xenobiotic-free synthetic cell culture matrix that fully supports maintenance and directed differentiation of human induced pluripotent stem cells (hiPSCs) down all three germ layers and dramatically reduces the costs of stem cell culture processes. The current, most commonly used matrices either create significant reliability and safety issues, or are effective only with certain cell lines given their limited mechanisms for cel adhesion. To address these drawbacks, Primorigen will develop a new synthetic cell culture matrix that recapitulates key attributes of natural extracellular matrix by presenting diverse synthetic ligands that offer numerous mechanisms for cell adhesion and facilitate sequestration of endogenous factors thereby reducing the need for and costs of using exogenous growth factors. Phase I studies will demonstrate feasibility by (a) creating a library of synthetic cell adhesion and endogenous factor sequestering ligands, (b) optimizing the coating conditions of ligand mixtures onto polystyrene surfaces, and (c) validating the new synthetic cell culture matrix for hiPSC maintenance and directed differentiation into various lineages. Phase II objectives will build on this work by expanding the synthetic ligand mixture approach to validate their use in large scale production and differentiation of hiPSCs. Commercial outputs will include (i) a broadly applicable synthetic Matrigel replacement comprised of synthetic modular ligand mixtures optimized for stem cell maintenance and differentiation, (ii) polystyrene tissue culture platforms (e.g., Petri dish, multiwell plate and microcarrier) pre-coated with the synthetic modular ligand mixtures, and (iii) a library of synthetic modular ligands from which made to order, customized mixtures can be assembled according to individual customers' unique needs.