Regulation of Growth and Differentiation in 3-D Cultures of Olfactory Epithelium

嗅上皮 3D 培养中生长和分化的调节

• 批准号: 8196734
• 负责人: JAMES E. SCHWOB
• 金额: $ 20.63万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-01 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8196734
• 关键词: 3-Dimensional; Adult; Affect; Air; Animals; Autologous; Biological Assay; Biological Markers; Biological Preservation; Birth; Cataloging; Catalogs; Cell Death; Cell Line; Cells; Chemistry; Complex; Conditioned Culture Media; Cues; Data; Differentiation and Growth; Dimensions; Dissociation; Epithelial; Epithelium; Exogenous Factors; Exploratory/Developmental Grant; Failure; Fibroblasts; Fostering; Generations; Goals; Growth; Growth Factor; Health; Image Analysis; In Vitro; Individual; Injury; Label; Lamina Propria; Lesion; Life; Maintenance; Measures; Metabolic; Modeling; Molecular; Mucous Membrane; Mus; Natural regeneration; Nature; Neonatal; Nervous System Part; Neuroglia; Neurons; Olfactory Epithelium; Olfactory Mucosa; Pathology; Patients; Play; Population; Process; Production; Property; Proteins; Regulation; Role; Signal Transduction; Smell Perception; Source; Spinal Cord; Stem cells; Stromal Cells; System; Testing; Therapeutic; Tissues; Translating; Transplantation; Work; analog; cell type; enhancing factor; exhaust; experimental analysis; genetic manipulation; in vitro Model; in vivo; injured; neurogenesis; prevent; protein profiling; public health relevance; reconstitution; repaired; research study; stem; tandem mass spectrometry; tissue culture; two-dimensional

DESCRIPTION (provided by applicant): The capacity for regeneration of neuronal and non-neuronal populations in the olfactory epithelium (OE) is well-known and extends throughout life. By virtue of their accessibility, the neurocompetent stem and progenitor cells of the OE are attractive candidates for use in autologous cellular therapies. The regulatory signals and mechanisms that govern these processes are not well understood. An in-depth analysis of the regulation of olfactory epitheliopoeisis is needed in order to exploit the stem and progenitor cells fully, but progress has been slow. We have developed a tissue culture assay in which cells from the neonatal or adult lesioned OE grow within spheres that form at the air-media interface of culture well inserts in a condition- dependent manner. Remarkably, cells that are cultivated in 3-dimensions (3-D) retain the potency and plasticity to participate in the regeneration of the OE in the animal, which stands in sharp contrast to the failure of cells grown in 2-D to do so. Two Aims are proposed to explore the cues regulating cell generation and differentiation taking advantage of the in vivo-like properties of the sphere assay. Aim 1 will test how growth factors, alone or in combination, will affect the growth of OE cells in the 3-D cultures. Some of the factors have a previously established role in the turnover and regeneration process in the OE; they will calibrate the assay, providing registration of the in vitro and in vivo results. For the majority, little is known of their role in the OE, although they function elsewhere in cell birth or differentiation. In contrast, Aim 2 will determine the relevant cues within a complex molecular mix that is known to play an important role in regulating the OE. In this case, culture media conditioned by the growth of a lamina propria cell line (derived from the part of the mucosa that is deep to the OE) is required for the formation of spheres from adult lesioned OE on inserts. We will take both a candidate molecule and an open-ended approach to identifying the relevant factors. PUBLIC HEALTH RELEVANCE: The health relevance of the work resides in the ultimate goal of achieving regulated controlled growth of olfactory stem cells and progenitor cells. Besides the goal of helping patients with dysosmias or anosmias, the neurocompetent tissue stem cells may prove useful for repairing other parts of the nervous system, including the spinal cord after it is injured.

描述（由申请人提供）：嗅觉上皮（OE）中神经元和非神经元种群再生的能力是众所周知的，并且遍及一生。凭借其可及性，OE的神经能力茎和祖细胞是可用于自体细胞疗法的有吸引力的候选者。管理这些过程的监管信号和机制尚不清楚。为了充分利用茎和祖细胞，需要对嗅觉上皮性的调节进行深入分析，但进展很慢。我们已经开发了一种组织培养测定法，其中来自新生儿或成年病变的OE的细胞在培养井媒体界面形成的球体内生长，并以条件依赖性方式插入。值得注意的是，在三维中培养的细胞（3-D）保留了参与动物中OE再生的效力和可塑性，与在2-D中生长的细胞失败形成鲜明对比。提出了两个目的，以探索利用球体测定的体内特性的调节细胞产生和分化的提示。 AIM 1将测试生长因子单独或组合如何影响3-D培养物中OE细胞的生长。其中一些因素在OE的离职和再生过程中具有先前确定的作用。他们将校准测定，提供体外和体内结果的注册。对于大多数人来说，尽管它们在细胞出生或分化方面的作用，但知之甚少。相比之下，AIM 2将确定复杂分子混合物中的相关线索，该提示在调节OE中起着重要作用。在这种情况下，培养基是由固有椎板细胞系的生长（源自粘膜深处到OE的一部分）的培养基，才能在插入物上形成成人病变的OE球体。我们将同时采用候选分子和开放式方法来识别相关因素。 公共卫生相关性：这项工作的健康相关性源于实现受管制的嗅觉干细胞和祖细胞受控生长的最终目标。除了帮助患有障碍症或厌氧的患者外，神经能力的组织干细胞可能被证明可用于修复神经系统的其他部位，包括脊髓受伤后的脊髓。