A Novel Defined Xenobiotic Free Substrate that Mimics Natural Extracellular Matrix

Primorigen Biosciences将开发一种明确的、不含外源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）在所有三个胚层的维持和定向分化，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显著的可靠性和安全性问题，要么只对某些细胞系有效，因为它们的细胞粘附机制有限。为了解决这些缺点，Primorigen将开发一种新的合成细胞培养基质，通过提供多种合成配体，提供多种细胞粘附机制，促进内源因子的隔离，从而减少使用外源生长因子的需求和成本，从而概括了天然细胞外基质的关键属性。I期研究将通过(a)创建一个合成细胞粘附和内源性因子隔离配体库，(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用于维持hiPSC并定向分化为各种世系来证明可行性。第二阶段的目标将建立在这项工作的基础上，扩大合成配体混合物的方法，以验证它们在大规模生产和分化hipsc中的应用。商业产出将包括(i)广泛适用的合成Matrigel替代品，由合成模块化配体混合物组成，优化干细胞维持和分化，（ii）预先涂覆合成模块化配体混合物的聚苯乙烯组织培养平台（例如，培养皿、多孔板和微载体），以及（iii）合成模块化配体库。可根据个别客户的独特需求组装定制混合物。

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.