Optimizing Assays of Human Stem Cells in Bone Marrow

优化人类骨髓干细胞的检测

• 批准号: 7094092
• 负责人: George F Muschler
• 金额: $ 37.35万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7094092
• 关键词: aging; bioassay; bone marrow; cell age; cell cycle; cell population study; clinical research; cytology; flow cytometry; hematopoiesis; hematopoietic stem cells; histogenesis; human subject; image processing; physiology; sample collection; southern blotting

DESCRIPTION (provided by applicant): Throughout life all tissues are maintained by underlying systems of stem cells that continually renew tissues through the generation of new Cells. These systems involve a series of hierarchical stages, originating with a rare population of upstream stem cells, and progressing through more numerous populations of downstream progenitor cells. Aging is associated with (and possibly defined by) changes in tissue health, appearance, and capacity for repair that are inevitably associated with underlying changes in the number of stem cells and/or their downstream kinetics (e.g. activation, self renewal, proliferation, migration, differentiation, and survival). Bone marrow is particularly relevant to aging. No tissue undergoes such profound change with age. Moreover, bone marrow is the home of multiple stem cell pools. Hematopoetic stem cells (HSCs) support the formation and maintenance of blood cells and the immune system. Mesenchymal Stem Cells (MSCs) and downstream connective tissue progenitors (CTPs), support the generation of bone, cartilage, fat, fibrous tissue, and skeletal muscle. Furthermore, even less numerous stem cell pools in bone marrow (e.g. multipotent adult progenitor cells (MAPCs), side population (SP) cells) have been shown to be capable of contributing to tissues of multiple germ layers, including liver, brain, lung and gut. This multidimensional nature places bone marrow in a position of central importance for understanding stem cell systems and the age related changes in these systems that contribute to preservation of health and functional accommodation to disease. However, conventional methods for harvest and sampling of bone marrow derived cells by aspiration is not well suited for quantitative assay of the stem cell population, due to large errors associated with sampling bias and dilution and contamination with cells from blood. Furthermore, aspiration disrupts information related to stem cell function that is provided by features of distribution and tissue level organization of stem cell and progenitor populations. This proposal will address deficiencies using an integrated cell-based modeling strategy to enable the systematic assessment of stem cell populations, and provide objective information necessary to optimally perform and interpret stem cell assays available from aspirate samples and small diameter bone core biopsies. 160 patients undergoing elective hip and knee arthroplasty (age 30 to 90), will provide the opportunity to assess bone and marrow samples from a broad age range. An integrated multidisciplinary analysis strategy will be applied to each sample, including: fluorescence activated cell sorting (FACS) and cell fraction analysis, CTP colony assay, LTClC assay, assay of telomerase and telomere length, quantitative histomorphometry of bone and marrow and pathological assessment.

描述（由申请人提供）：在整个生命中，所有组织都通过干细胞的潜在系统来维持，这些干细胞的潜在系统通过新细胞的产生不断更新组织。这些系统涉及一系列分层阶段，起源于罕见的上游干细胞，并在许多下游祖细胞中发展。衰老与组织健康，外观和修复能力的变化有关，这些变化与干细胞数量和/或其下游动力学的潜在变化（例如激活，自我更新，增生，增殖，迁移，差异，差异和存活）不可避免地相关。 骨髓与衰老特别相关。随着年龄的增长，没有组织会发生如此深刻的变化。 此外，骨髓是多个干细胞池的所在地。造血干细胞（HSC）支持血细胞和免疫系统的形成和维持。间充质干细胞（MSC）和下游结缔组织祖细胞（CTP），支持骨，软骨，脂肪，纤维组织和骨骼肌的产生。此外，骨髓中的干细胞池甚至较少的干细胞池（例如，多能成年祖细胞（MAPC），侧种群（SP）细胞）被证明能够为包括肝脏，脑，肺和肠道在内的多个细菌层的组织做出贡献。这种多维性质使骨髓处于理解干细胞系统以及这些系统中与年龄相关的变化有助于保存健康和疾病功能适应的重要性。然而，由于与血液中的采样偏见，稀释和污染细胞有关的较大误差，通过抽吸衍生细胞收集和采样骨髓衍生细胞的常规方法不适合对干细胞种群进行定量测定。此外，抽吸会破坏与干细胞功能有关的信息，该信息由干细胞和祖细胞种群的分布和组织水平组织的特征提供。 该建议将使用基于整合的细胞建模策略来解决缺陷，以实现干细胞群体的系统评估，并提供必要的客观信息，以最佳地执行和解释从抽吸样品和小直径骨核心活检中获得的干细胞测定。 160例接受选长髋关节和膝关节置换术的患者（30至90岁）将为评估广泛年龄范围的骨骼和骨髓样品提供机会。每个样本将应用一个综合的多学科分析策略，包括：荧光激活的细胞分选（FACS）和细胞分数分析，CTP菌落分析，LTCLC测定，端粒酶和端粒长度测定，骨骼和骨髓的定量组织形态学和病理评估。