Functional dissection of human adipose and bone marrow-derived MSC phenotypes

人类脂肪和骨髓来源的 MSC 表型的功能解剖

• 批准号: 7653541
• 负责人: Jan A. Nolta
• 金额: $ 38.25万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-20 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7653541
• 关键词: Acute; Adipose tissue; Adverse event; Applications Grants; Area; Blood; Blood Circulation; Bone Marrow; CXCR4 gene; Cartilage; Cell Culture Techniques; Cell Surface Receptors; Cell surface; Cells; Clinical; Clinical Trials; Clone Cells; Data; Dissection; Environment; FDA approved; Fatty acid glycerol esters; Funding; Goals; Grant; Health Care Costs; Healthcare; Hindlimb; Home environment; Homing; Human; Human Characteristics; Hypoxia; Immune; Immunohistochemistry; In Situ Hybridization; In Vitro; Individual; Inflammatory; Injection of therapeutic agent; Injury; Integrins; Inverse Polymerase Chain Reaction; Ischemia; Laboratories; Learning; Ligand Binding; Limb structure; Liver; Marrow; Mediating; Membrane Proteins; Mesenchymal Stem Cells; Methods; Modeling; Molecular; Mus; Muscle; Outcome; Oxygen; Paramedical Personnel; Patients; Phenotype; Physiological; Population; Proteins; Publishing; Reproducibility; Signal Transduction; Small Interfering RNA; Sorting - Cell Movement; Stem cells; Stromal Cell-Derived Factor 1; Techniques; Testing; Tissues; Uncertainty; Viral Vector; Work; Wound Healing; Xenograft procedure; adult stem cell; aldehyde dehydrogenases; base; bone; candidate marker; clinically relevant; effective therapy; improved; in vivo; injured; interest; knock-down; meetings; migration; novel; paracrine; prospective; receptor; receptor expression; repaired; research study; self-renewal; stem cell biology; stem cell population; tool; trafficking; wound

We have characterized the in vitro and in vivo characteristics of human adipose-derived mesenchymal stem cells (A-MSC), and compared them to marrow MSC (BM-MSC). We learned that MSC lodge at low levels into multiple tissues in immune deficient mice, but that both BM-MSC and A-MSC migrate from the bloodstream into areas of hypoxic tissue damage much more robustly, and we are continuing to define the mechanisms involved. We have dissected putative markers and phenotypes, and learned that surface proteins are altered dependent upon microenvironmental factors such as oxygen concentration and ligand binding. Our hypothesis is that the “tissue-repairing subset” of MSC is composed of highly adaptable cells that are poised to respond rapidly to the wound or inflammatory microenvironment by altering cell surface receptor expression, integrins, MMPS, and other proteins related to migration, retention, and survival. MSC are “paramedics”, navigating through tissue to secrete paracrine factors that initiate cascades of endogenous repair and revascularization. It is beneficial if they are able to respond to the environment and to adapt very rapidly. But, this quality makes it difficult to prospectively isolate MSC for tissue repair. Thus, the phenotype of the most primitive MSC compartment, to allow prospective isolation from marrow or adipose tissue, is still not well defined and remains a goal of the grant application. We hypothesize that standard MSC culture conditions, which are non-physiological, are differentiating MSC to states that are not compatible with maximal tissue repair capacity. The goal of the current grant, reduced in scope is to subfractionate and to continue to examine MSC phenotype, in standard culture and in more physiological conditions, with the goal of prospectively isolating and potentially expanding primitive populations which retain the ability to form cartilage, bone, and fat and/or to home to damaged tissues. We will compare the phenotype and function of MSC expanded under standard conditions to more physiologic conditions, in comparison to prospectively isolated ALDH hi/CD45-/CD31-MSC populations. We will use clonal integration marking and novel xenotransplantation models to define conditions that allow recruitment of the highest numbers of primitive human MSC to areas of tissue damage, and to better define multipotency and self-renewal in this interesting adult stem cell population.

我们已经表征了人脂肪间充质干细胞（A-MSC）的体外和体内特性，并将其与骨髓MSC（BM-MSC）进行比较。我们了解到MSC在免疫缺陷小鼠中以低水平进入多个组织，但BM-MSC和A-MSC都从血流中迁移到缺氧组织损伤区域，并且我们正在继续确定所涉及的机制。我们已经剖析了推定的标记物和表型，并了解到表面蛋白质的改变取决于氧浓度和配体结合等微环境因素。我们的假设是，MSC的“组织修复亚群”由高度适应性的细胞组成，这些细胞通过改变细胞表面受体表达、整合素、MMPS和其他与迁移、滞留和存活相关的蛋白质，对伤口或炎症微环境做出快速反应。MSC是“护理人员”，在组织中导航以分泌旁分泌因子，启动内源性修复和血管重建的级联反应。如果他们能够对环境做出反应并迅速适应，这是有益的。但是，这种性质使得难以前瞻性地分离MSC用于组织修复。因此，允许从骨髓或脂肪组织中前瞻性分离的最原始MSC隔室的表型仍然没有很好地定义，并且仍然是资助申请的目标。我们假设，标准的MSC培养条件，这是非生理，分化MSC的状态，是不兼容的最大组织修复能力。目前资助的目标是在标准培养和更多的生理条件下进行亚骨折并继续检查MSC表型，目标是前瞻性地分离和潜在地扩大保留形成软骨，骨和脂肪和/或修复受损组织的能力的原始群体。我们将比较在标准条件下扩增的MSC的表型和功能，与前瞻性分离的ALDH hi/CD 45-/CD 31-MSC群体进行比较。我们将使用克隆整合标记和新的异种移植模型来定义允许招募最高数量的原始人MSC到组织损伤区域的条件，并更好地定义这个有趣的成体干细胞群体的多能性和自我更新。