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.

抽象t 脂肪组织来源的间充质干细胞（AD-MSCs）有潜力作为一个重要的工具， 对于骨组织工程来说，因为它们在体内相对较高的频率可以提供极大的 再生疗法所需的细胞数量。不幸的是，许多研究表明，AD-MSCs 与其他类型的干细胞相比，成骨潜力要小得多。我们的团队已经开发出一种 体外产生的细胞外基质（ECM），重现了天然骨髓（BM-）的特定属性 脂肪微环境（AD-）在以前的研究中，我们证明了BM-ECM显著地 相对于标准培养塑料，促进骨髓（BM）来源的MSC的成骨分化。在 在目前的建议中，我们评估了BM-ECM增加AD-MSC成骨能力的潜力， 并且通过这样做，显著地提高它们对于骨骼修复和再生的临床相关性。 我们的初步数据表明BM和AD来源的ECM能够指导BM-MSC分化 向成骨细胞和脂肪细胞谱系发展。这些ECM的特点是在以下方面存在显著差异： 生化成分、结构甚至机械性能。我们证明了BM-和AD- 在两种组织特异性ECM上培养的MSC更容易增殖，当每种类型的MSC都是 维持在源自相同的相应组织的ECM上。此外，我们观察到BM-和AD- MSCs在BM-ECM上均呈低圆度铺展，而在AD-ECM上呈高圆度铺展 扩展形态学这些观察结果与最近的报告相一致，这些报告表明， MSC形态最终与涉及谱系决定的信号线索有关。此外，最近 研究表明，粘着斑复合物的形成可以调节干细胞的命运决定， 经典的Wnt/β-catenin通路。我们将研究是否在组织上形成独特的粘着斑复合物- 特异性ECM，并重建负责指导MSC分化的微环境， 成骨细胞和脂肪细胞谱系。 作为这些观察的结果，我们假设BM-ECM的骨特异性微环境 可以“再训练”AD-MSC以增强其成骨潜力。具体目标如下： 1）为了确定BM-和AD-MSC分化的差异，评估 用体外和体内成骨和脂肪生成试验，用组织培养塑料（TCP）培养 和组织特异性ECM，以及2）确定负责促进MSC 通过在TCP和组织特异性ECM上培养，可诱导成骨细胞和脂肪细胞谱系的分化。