MATRIX, HORMONES, AND DIFFERENTIATION IN MAMMALIAN CELLS

长期目标是建立示范系统和文化 分析生长调节机制的条件 和/或正常和肿瘤上皮细胞的分化。 的 正在开发的模型系统是前列腺上皮细胞和正常的 肝细胞 该方法是基于这样的假设，即增长和 上皮细胞的分化受到复杂的细胞因子的影响。 存在于组织中的相互作用，并且可以在 培养条件 我们所关注的细胞相互作用是 上皮-间充质关系，其可被刺激， 基底膜的利用，细胞外基质的一种形式， 特定的激素和生长因子，其混合物被设计用于 每种特定的细胞类型。 我们的目标是继续发展 适当的底物和确定的培养基条件， 维持正常前列腺上皮细胞和正常前列腺上皮细胞的培养 处于生长状态或最大表达状态的肝细胞， 它们的组织特异性功能 到目前为止，我们已经开发了用于 分离组织特异性形式的细胞外基质， 用作培养细胞的底物。 将细胞接种到这些 基质基底显示出增强的附着和电镀效率 并且生长或分化反应更类似于它们在体内的反应 应答 结合这种方法，我们正在利用 佐藤和他的同事开发“定义媒体”的方法， 为特定细胞设计的无血清、经血液补充的培养基 类型 我们发现，基质的类型和基质的组成 成分确定的培养基不仅随细胞类型而变化， 细胞的生理反应。 因此，基质和成分确定培养基 特定细胞类型生长所需的细胞数量与所需的细胞数量不同。 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， 在这些条件下维持的细胞培养物的表征 条件允许我们最终以系统的方式改变， 生长调节和调节分析中的条件 分化

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.