Regenerative acellular biomaterials derived from embryonic stem cells

描述（由申请人提供）：在大多数成年哺乳动物组织中，瘢痕组织形成是对损伤的常见反应，它会严重损害受损组织的完整性和功能。用于组织修复的再生细胞外基质（ecm）由天然生物分子组成，可以通过脱细胞化方法从各种细胞和组织来源中获得。胚胎干细胞是最原始的干细胞，能够分化为所有体细胞类型，包括三种生殖谱系（外胚层、内胚层和中胚层）。胚胎干细胞通过胚状体（EB）的形成在体外自发分化，其中悬浮中形成的多细胞聚集体再现了组织形态发生前原肠胚形成阶段调节细胞信号传导和命运决定的分子微环境。我们已经开发了一种脱细胞化技术，从经过EB形成分化的ES细胞中产生新的细胞外基质。本研究的目标是：1)用不同的溶剂提取技术确定EB脱细胞化的效率；2)量化所得脱细胞材料中ECM分子的保留率；3)在小鼠皮肤伤口愈合模型中评估EB细胞衍生的ECM的功能再生潜力。这些目标的完成将为胚胎干细胞在早期分化过程中产生的原始ECM的组成和功能提供新的生物学信息。此外，这些研究将在成人损伤模型中检测直接来源于胚胎干细胞的新型脱细胞胚胎ECM的再生能力。这种方法代表了将干细胞，特别是干细胞衍生材料应用于再生医学治疗的全新概念。在成年哺乳动物中，瘢痕组织的形成是由于对大多数损伤和许多慢性疾病的伤口愈合反应不足而导致的一个重要问题。

DESCRIPTION (provided by applicant): Scar tissue formation is a common response to injury in most tissues of adult mammals and it can significantly impair the integrity and function of afflicted tissues. Regenerative extracellular matrices (ECMs) for tissue repair, comprised of natural biological molecules, can be derived from various cell and tissue sources via acellularization approaches. Embryonic stem (ES) cells are the most primitive stem cells capable of differentiating into all somatic cell types comprising the three germ lineages (ectoderm, endoderm and mesoderm). ES cells spontaneously differentiate in vitro via embryoid body (EB) formation, whereby multi-cellular aggregates formed in suspension recapitulate the molecular microenvironment regulating cell signaling and fate decisions in the pre- gastrulation stages of tissue morphogenesis. We have developed an acellularization technique to yield novel extracellular matrices from ES cells undergoing differentiation via EB formation. The objectives of this proposal are to 1) determine the efficiency of acellularization of EBs with different solvent extraction techniques, 2) quantify the retention of ECM molecules within the resulting acellular material and 3) assess the functional regenerative potential of the ECM derived from EB cells in a murine dermal wound healing model. The completion of these aims will yield new biological information about the composition and function of primitive ECM produced by ES cells during the course of early differentiation. In addition, these studies will examine the regenerative capability of a novel acellular embryonic ECM derived directly from ES cells in an adult injury model. This approach represents a fundamentally new concept for the application of stem cells, and more specifically stem cell-derived materials, to regenerative medicine therapies. Scar tissue formation is a significant problem that results from an inadequate wound healing response to most injuries and many chronic diseases in adult mammalian species.