Tissue-Specific Extracellular Matrix for Culturing Stem and Progenitor Cells

性状（申请人提供）：不同组织的细胞外基质不同。由于正常的健康细胞创造了它们自己的最佳环境，因此当这些细胞被衍生时，可以从组织中创造用于细胞培养的最佳环境。令人惊讶的是，目前没有这样的“培养皿”可供有兴趣研究组织特异性再生细胞类型（所谓的成体干细胞）的生长和分化的研究人员使用。虽然有通用的涂层和3-D支架，但这些产品并不特定于许多研究人员正在研究的各种成年哺乳动物干细胞和祖细胞，因此它们并不适合给定的系统。一些研究人员最近表明，细胞外基质（ECM）成分的细微变化可以对培养物中的生长，分化和组织形成产生显着影响。为此，我们建议研究提供细胞-基质相互作用的组织特异性ECM涂层，以及这些相互作用可能对成体干细胞和祖细胞的生长和分化产生的影响。通过这样做，我们将为哺乳动物细胞，特别是干细胞和祖细胞的体外生长和分化提供必要的工具，从而加快再生医学许多领域的发展步伐。在这项研究中，我们将首先致力于开发细胞培养涂层，以证明我们的方法的可行性。我们将在作为目标基质的3种相关ECM涂层上测试3种细胞类型：1）分化为角质形成细胞的皮肤来源干细胞（无细胞真皮ECM），2）分化为骨骼肌成肌细胞的肌肉卫星细胞（骨骼肌ECM），以及3）分化为肝细胞的羊水来源干细胞（肝脏ECM）。这将通过使用我们的组织去细胞化的标准方案，通过开发脱细胞组织消化的新方案，并最终通过将这些包被物涂覆到组织培养皿上来实现。将检测细胞在其各自基质以及其他基质上的生长和分化潜力，以证明ECM-细胞相互作用的特异性。表征将包括定量（例如流式细胞术）定性（例如免疫细胞化学）和功能（即代谢）测定。我们推测，由组织特异性ECM介导的细胞-基质相互作用将促进更有效、完整和快速的分化，从而导致更多功能的组织再生。这些新的生物材料最终可能会导致基于细胞和组织的疗法的更快速发展，以及更有效的工业规模扩大和临床转化。

DESCRIPTION (provided by applicant): Extracellular matrix from different tissues is different. Since normal healthy cells create their own optimal environment, it follows that an optimal environment for cell culture can be created from the tissue when these cells were derived. Surprisingly, no such "cultureware" is currently available to researchers interested in investigating the growth and differentiation of tissue-specific regenerative cell types (so called adult stem cells). While there are generic coatings and 3-D scaffolds, these products are not specific to the various adult mammalian stem and progenitor cells being studied by many investigators, hence they are not optimized for a given system. Several researchers have recently shown that subtle changes to extracellular matrix (ECM) components can have a dramatic effect on growth, differentiation, and tissue formation in culture. To that end, we propose to study tissue-specific ECM coatings that provide cell-substrate interactions, and the effect these interactions may have on growth and differentiation of adult stem and progenitor cells. In so doing, we will provide essential tools for the in vitro growth and differentiation of mammalian cells, specifically stem and progenitor cells, thereby accelerating the pace of development in many areas of regenerative medicine. In this study, we will first aim to develop cell culture coatings to demonstrate the feasibility of our approach. We will test 3 cell types on 3 related ECM coatings as the substrate of interest: 1) skin-derived stem cells differentiated into keratinocytes (acellular dermis ECM), 2) muscle satellite cells differentiated into skeletal myoblasts (skeletal muscle ECM), and 3) amniotic fluid-derived stem cells differentiated into hepatocytes (liver ECM). This will be accomplished by using our standard protocols for tissue decellularization, by developing new protocols for acellular tissue digestion, and finally by coating these digestates onto tissue culture dishes. Cells will be tested for growth and differentiation potential on their respective substrates, as well as the other substrates, to demonstrate the specificity of the ECM-cell interactions. Characterization will include both quantitative (e.g. flow cytometry) qualitative (e.g. immunocytochemistry), and functional (i.e. metabolic) assays. We hypothesize that the cell-substrate interactions mediated by tissue-specific ECM will facilitate more efficient, complete, and rapid differentiation, thereby resulting in more functional tissue regeneration. These new biomaterials could eventually lead to more rapid development of cell and tissue-based therapies, as well as more efficient industrial scale-up and clinical translation.