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群体的再生能力是众所周知的，并贯穿整个生命。由于其可获得性，OE的神经功能干细胞和祖细胞是用于自体细胞治疗的有吸引力的候选细胞。管理这些过程的监管信号和机制还没有得到很好的理解。为了充分利用干细胞和祖细胞，需要对嗅觉上皮细胞的调控进行深入的分析，但进展缓慢。我们已经开发了一种组织培养试验，其中来自新生儿或成人受损OE的细胞以条件依赖的方式在培养井的空气-介质界面形成的球体内生长。值得注意的是，在三维(3-D)培养的细胞保持了参与动物OE再生的能力和可塑性，这与在2-D培养的细胞无法做到这一点形成了鲜明的对比。利用球体实验的体内相似特性，提出了两个目的来探索调控细胞生成和分化的线索。目标1将测试生长因素如何单独或联合影响3-D培养中的OE细胞的生长。其中一些因素在OE的周转和再生过程中具有先前确立的作用；它们将校准检测，提供体外和体内结果的注册。对于大多数人来说，人们对它们在OE中的作用知之甚少，尽管它们在细胞出生或分化的其他地方发挥作用。相反，目标2将确定复杂分子混合物中的相关线索，该复杂分子混合物已知在调节OE方面发挥重要作用。在这种情况下，需要以固有层细胞系(来自OE深处的黏膜部分)的生长为条件的培养液，以形成插入件上的成人受损OE球体。我们将采用候选分子和开放式方法来确定相关因素。 公共卫生相关性：这项工作的健康相关性在于实现嗅觉干细胞和祖细胞受控生长的最终目标。除了帮助嗅觉障碍或嗅觉障碍患者的目标外，具有神经功能的组织干细胞可能被证明对修复神经系统的其他部分有用，包括损伤后的脊髓。