Regenerative acellular biomaterials derived from embryonic stem cells

描述（由申请人提供）：瘢痕组织形成是成年哺乳动物大多数组织中对损伤的常见反应，并且其可显著损害受累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获得。胚胎干细胞（ES）是能够分化成包括三个生殖谱系（外胚层、内胚层和中胚层）的所有体细胞类型的最原始的干细胞。ES细胞通过胚状体（EB）形成在体外自发分化，由此在悬浮液中形成的多细胞聚集体概括了在组织形态发生的原肠胚形成前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去细胞化技术，以产生新的细胞外基质的ES细胞通过EB形成分化。该提议的目的是1）确定用不同溶剂提取技术使EB去细胞化的效率，2）量化ECM分子在所得去细胞材料内的保留，和3）评估源自小鼠皮肤伤口愈合模型中EB细胞的ECM的功能再生潜力。这些目标的完成将产生新的生物学信息的组成和功能的原始ECM产生的ES细胞在早期分化过程中。此外，这些研究将检查一种新的脱细胞胚胎ECM的再生能力，直接从ES细胞在成人损伤模型。这种方法代表了干细胞，更具体地说是干细胞衍生材料应用于再生医学治疗的一个全新概念。瘢痕组织形成是一个重要的问题，它是由成年哺乳动物物种对大多数损伤和许多慢性疾病的伤口愈合反应不足引起的。

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.