Control of Stem Cell Behavior by Exposure to Tissue-Specific ECM

通过暴露于组织特异性 ECM 来控制干细胞行为

• 批准号: 9259665
• 负责人: Milos Marinkovic
• 金额: $ 3.82万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9259665
• 关键词: Address; Adipocytes; Adipose tissue; Age; Architecture; Behavior; Biochemical; Biological Assay; Blood Vessels; Bone Marrow; Bone Marrow Stem Cell; Bone Tissue; Cell Count; Cell Culture Techniques; Cell Lineage; Cell Therapy; Cells; Chondrocytes; Clinical; Collection; Complex; Cues; Data; Development; Down-Regulation; Education; Exhibits; Exposure to; Extracellular Matrix; Fatty acid glycerol esters; Figs - dietary; Focal Adhesions; Frequencies; Heterogeneity; Histocompatibility Testing; In Vitro; Inferior; Knowledge; Mechanics; Mesenchymal Stem Cells; Modeling; Molecular; Mononuclear; Morbidity - disease rate; Morphology; Multipotent Stem Cells; Myoblasts; Natural regeneration; Osteoblasts; Osteogenesis; Pathway interactions; Population; Proliferating; Property; Regenerative Medicine; Reporting; Research Personnel; Signal Transduction; Stem cells; Stromal Cells; Suction Lipectomy; Testing; Tissue Engineering; Tissues; Training; Translations; Up-Regulation; Work; abstracting; adipocyte differentiation; base; beta catenin; bone; cell behavior; clinically relevant; improved; in vivo; innovation; lipid biosynthesis; osteogenic; reconstitution; regenerative therapy; repaired; skeletal; skeletal regeneration; skeletal tissue; stem cell differentiation; stem cell fate; stem cell niche; stemness; tissue culture; tool

Abstract t Adipose tissue-derived mesenchymal stem cells (AD-MSCs) have the potential to serve as a critical tool for bone tissue engineering as their relatively high frequency in the body can provide the exceedingly large numbers of cells required for regenerative therapies. Unfortunately, many studies have shown that AD-MSCs have considerably less osteogenic potential than other types of stem cells. Our group has developed an in vitro-produced extracellular matrix (ECM) that recapitulates specific attributes of the native bone marrow (BM-) and adipose (AD-) microenvironment. In previous studies, we demonstrated that BM-ECM significantly promotes osteogenic differentiation by bone marrow (BM)-derived MSCs relative to standard culture plastic. In the current proposal, we evaluate the potential of BM-ECM to increase the osteogenic capacity of AD-MSCs, and by doing so, dramatically improve their clinical relevance for skeletal repair and regeneration. Our preliminary data show that BM- and AD-derived ECMs are capable of guiding BM-MSC differentiation towards osteoblast and adipocyte lineages. These ECMs are characterized by significant differences in biochemical composition, architecture and even mechanical properties. We demonstrated that BM- and AD- MSCs cultured on both types of tissue-specific ECM proliferated more readily when each type of MSC was maintained on ECM-derived from the same respective tissue. Additionally, we observed that BM- and AD- MSCs both exhibited low-circularity spreading on BM-ECM, while on AD-ECM, they displayed a high-circularity spreading morphology. These observations are consistent with recent reports demonstrating that changes in MSC morphology are ultimately related to signaling cues involved in lineage-decisions. Furthermore, recent studies have suggested that focal adhesion complex formation can regulate stem cell fate decisions through the canonical Wnt/β-catenin pathway. We will investigate if unique focal adhesion complexes form on tissue- specific ECMs and reconstitute the microenvironment(s) responsible for directing MSC differentiation to either the osteoblast and adipocyte lineage. As a result of these observations, we hypothesize that the bone-specific microenvironment of BM-ECM can "re-train" AD-MSCs in order to enhance their osteogenic potential. The following specific aims are proposed to test this hypothesis: 1) To determine differences in BM- and AD-MSC differentiation, assessed with assays of osteogenesis and adipogenesis in vitro and in vivo, with culture on tissue culture plastic (TCP) and tissue-specific ECM and 2) To determine the molecular mechanism(s) responsible for promoting MSC differentiation to the osteoblast and adipocyte lineages with culture on TCP and tissue-specific ECM.

摘要t 脂肪组织衍生的间充质干细胞（AD-MSC）具有作为关键工具的潜力 对于骨组织工程，由于其体内的相对高频可以提供更大的大型 再生疗法所需的细胞数量。不幸的是，许多研究表明AD-MSCS 比其他类型的干细胞具有小心的成骨潜力。我们的小组发展了 体外生产的细胞外基质（ECM）概括了天然骨髓（BM-）的特定属性 和脂肪（AD-）微环境。在先前的研究中，我们证明了BM-ECM显着 相对于标准培养塑料，通过骨髓（BM）衍生的MSC促进成骨分化。在 当前的建议，我们评估了BM-ECM增加AD-MSC的成骨能力的潜力， 通过这样做，可以极大地提高其在骨骼修复和再生方面的临床意义。 我们的初步数据表明，BM和AD衍生的ECM能够指导BM-MSC差异化 朝着成骨细胞和脂肪细胞谱系。这些ECM的特征是 生化组成，结构甚至机械性能。我们证明了BM和AD- 在两种类型的组织特异性ECM上培养的MSC在每种类型的MSC均更容易增殖 维持在来自相同的相对组织的ECM衍生上。此外，我们观察到BM和AD- MSC均暴露于BM-ECM上的低圆形性，而在AD-ECM上，它们显示出高圆形性 传播形态。这些观察结果与最近的报告一致，表明变化 MSC形态最终与谱系决策中有关的信号提示有关。此外，最近 研究表明，焦点粘合剂复合物的形成可以通过 规范的Wnt/β-catenin途径。我们将研究组织上是否形成独特的粘着粘合剂复合物 特定的ECM并重建负责将MSC差异化为两者的微环境 成骨细胞和脂肪细胞谱系。 由于这些观察结果，我们假设BM-ECM的骨特异性微环境 可以“重新训练” AD-MSC，以增强其成骨潜力。以下具体目的是 提议检验以下假设：1）确定BM-和AD-MSC分化的差异，评估 通过在体外和体内进行成骨和脂肪形成的测定，并在组织培养塑料（TCP）上进行培养 和组织特异性ECM和2）确定负责促进MSC的分子机制 与TCP和组织特异性ECM上培养的成骨细胞和脂肪细胞谱系的区分。