MAST CELL DIFFERENTIATION IN VITRO

肥大细胞体外分化

• 批准号: 3138963
• 负责人: THOMAS F HUFF
• 金额: $ 9.39万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-02-01 至 1993-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3138963
• 关键词: Nippostrongylus; RNA; antibody receptor; carboxypeptidase; cell differentiation; chemical binding; chymosin; colony stimulating factor; complementary DNA; connective tissue; embryo /fetus cell /tissue; gene expression; genetic translation; glycosylation; growth factor; growth factor receptors; heparin; immunoglobulin E; interferons; interleukin 3; laboratory mouse; laboratory rabbit; laboratory rat; lymphatic tissue; mast cell; skin; tissue /cell culture

The mast cell is an attractive model system for studies of cellular differentiation because the phenotypic changes from progenitor cell to mast cell are great and the signals required to induce differentiation appear to be few as compared with other cells such as B lymphocytes. We have defined a late, committed stage in the differentiation of the mast cell progenitor just prior to granulation. This mast cell-committed progenitor does not require IL-3 for differentiation but does require a factor or factors provided by a connective tissue microenvironment such as murine embryonic skin. If the mast cell-committed progenitor can be successfully isolated and the connective tissue factors identified, a well-defined in vitro differentiation system could be developed to individually examine discrete changes that account for the mast cell phenotype. First, we propose to develop a new in vitro differentiation system whereby any of several mast cell features can be studied by following the response of late-stage progenitor cells to defined factors. For the goal to be realized, the following strategy will be used: We will obtain a purified preparation of mast cell- committed progenitors based on affinity for embryonic skin monolayers. The phenotype of these progenitors will be determined. We will precisely identify the factor or factors in monolayers of murine embryonic skin which are required for differentiation of the mast cell committed progenitor into mast cells. When this is done, the isolated progenitor cells will be triggered with purified preparations of the required factor to examine changes in activation events, expression of high-affinity IgE receptors and growth factor receptors, and the ability to dedifferentiate. Second, we propose to use the mast cell-committed progenitor culture system described in the grant proposal to determine if there are any known IgE regulatory factors which also affect mast cell differentiation. Third, we propose to adapt this differentiation system for rat studies in order to detect chymase, carboxypeptidase A, or heparin proteoglycan in mast cells derived from culture on embryonic skin monolayers. This will allow us to study mast cell heterogeneity (mucosal vs connective tissue type in an in vitro system.

肥大细胞是细胞研究的一个有吸引力的模型系统 分化是因为祖细胞的表型发生了变化 细胞到肥大细胞很大，并且诱导所需的信号 与其他细胞相比，分化似乎很少 比如B淋巴细胞。 我们已经定义了一个后期的、承诺的阶段 在肥大细胞祖细胞的分化之前 造粒。 这种肥大细胞定向祖细胞不 需要 IL-3 进行分化，但确实需要一个因子或 由结缔组织微环境提供的因素，例如 小鼠胚胎皮肤。 如果肥大细胞定向祖细胞 可以成功分离结缔组织因子 确定后，可以建立一个明确的体外分化系统 开发用于单独检查离散变化 对于肥大细胞表型。 首先，我们建议开发一种新的体外分化系统 由此可以通过以下方式研究任何几种肥大细胞特征 遵循晚期祖细胞对定义的反应 因素。 为了实现目标，将采取以下策略 使用：我们将获得肥大细胞的纯化制剂- 基于对胚胎皮肤的亲和力的定向祖细胞 单层。 这些祖细胞的表型将是 决定。 我们将准确识别其中的一个或多个因素 小鼠胚胎皮肤的单层细胞是必需的 肥大细胞定向祖细胞分化为肥大 细胞。 完成此操作后，分离的祖细胞将被 用所需因子的纯化制剂触发 检查激活事件的变化，高亲和力的表达 IgE 受体和生长因子受体，以及 去分化。 其次，我们建议使用肥大细胞定向祖细胞 拨款提案中描述的文化系统，以确定是否 有任何已知的 IgE 调节因素也会影响肥大 细胞分化。 第三，我们建议将这种分化系统应用于大鼠 检测食糜酶、羧肽酶 A 或 培养物中肥大细胞中的肝素蛋白多糖 胚胎皮肤单层。 这将使我们能够研究肥大细胞 异质性（体外实验中的粘膜与结缔组织类型 系统。