MATRIX, HORMONES, AND DIFFERENTIATION IN MAMMALIAN CELLS

长期目标是建立模范系统和文化 分析增长调控机制的条件 和/或正常和肿瘤上皮细胞的分化。这个 正在开发的模型系统是前列腺上皮细胞和正常 肝细胞。这种方法是基于这样的假设，即增长和 复杂细胞对上皮细胞分化的影响 存在于组织中的相互作用，可以分析和模拟 培养条件。我们关注的细胞相互作用是 可被刺激的上皮-间充质关系 基底膜的利用，细胞外基质的一种形式，以及 特定的激素和生长因子，它们的混合物是为 每种特定的细胞类型。我们的目的是继续发展 适当的基质和确定的介质条件，以允许 正常前列腺上皮细胞和正常前列腺上皮细胞培养中的维持 处于生长状态或最高表达状态的肝细胞 它们特定于组织的功能。到目前为止，我们已经开发出了 分离组织特定形式的细胞外基质，可以 用作培养细胞的底物。细胞被电镀到这些上 基质衬底显示出更高的附着力和电镀效率 生长或分化反应更类似于它们在体内的反应 回应。与这种方法相结合，我们利用 佐藤和他的同事们开发“定义媒体”的方法论， 为特定细胞设计的无血清、补充激素的培养液 类型。我们发现矩阵的类型和元素的组成 定义的介质不仅随单元类型而异，而且还随 细胞的生理反应。因此，矩阵和定义的介质 特定细胞类型的生长所需的与所需的不同 以维持细胞的最大分化。我们希望 完成这些培养条件的特征化和 在每一种情况下维持的细胞培养的特征 允许我们最终以系统的方式变化的条件 生长调控分析中的条件和调控 差异化。

The long-term objectives are to establish model systems and culture conditions by which to analyze the mechanisms of regulation of growth and/or differentiation of normal and neoplastic epithelial cells. The model systems being developed are prostatic epithelial cells and of normal hepatocytes. The approach is based on the hypothesis that the growth and differentiation of epithelial cells are effected by complex cellular interactions present in tissues and which can be analyzed and simulate in culture conditions. The cellular interaction on which we are focused is the epithelial-mesenchymal relationship which can be stimulated by utilization of basement membranes, a form of extracellular matrix, and of specific hormones and growth factors, the mixture of which is designed for each specific cell type. Our intent is to continue the development of appropriate substrate and defined medium conditions by which to permit the maintenance in culture of normal prostatic epithelial cells and of normal hepatocytes in either a state of growth or a state of maximal expression of their tissue-specific functions. To date, we have developed methods for the isolation of tissue-specific forms of extracellular matrix which can be utilized as substrates for cells in culture. Cells plated onto these matrix substrates have shown enhanced attachment and plating efficiencies and growth or differentiative responses more similar to their in vivo responses. In combination with this approach, we are utilizing the methodologies of Sato and his associates to develop "defined media", serum-free, hormonally supplemented media designed for specific cell types. We are finding that the types of matrix and the composition of the defined media vary not only with the cell type but also with the physiological response of the cells. Thus, the matrix and defined medium needed for growth of a particular cell type is distinct from that needed for the maintenance of the cell in maximal differentiation. We hope to complete the chaacterization of these culture conditions and the characterization of the cultures of cells maintained under each of these conditions permitting us ultimately to vary in a systematic way the conditions in an analysis of growth regulation and regulation of differentiation.