A higher immune system model by combination of several types of artificial tissues (connective tissue, carcinoma tissue, and lymphoid tissue)

本研究重点结合结缔组织、癌组织、淋巴组织、血管等几种人工组织构建“高等体外免疫系统模型”。为实施研究计划，“肿瘤免疫学学部”与“医学技术学部”联合组建了一个新的研究领域“肿瘤技术”。本研究期间获得的数据如下：(1)人工结缔组织的建立：我们在i型胶原凝胶中重建了由人成纤维细胞和人脐静脉源性内皮细胞（HUVEC）组成的体外结缔组织模型。在该模型中，HUVEC在不添加任何特殊生长因子的情况下迁移形成细血管网。(2)人工癌组织的发展：我们重建了两种类型的人工癌组织。一种是软组织类型，我们可以在显微镜下实时观察两种细胞类型（c…更多的癌细胞和成纤维细胞）的生长超过30天。另一种是准备用于免疫组织化学分析的硬组织类型。在这个模型中，我们可以通过联合使用细胞因子来控制癌细胞的生长模式，即扩张性生长或侵袭性生长。(3)人工淋巴组织的发育：人工淋巴组织组织在I型胶原网络中，辅以OKT-3和纤维蛋白原、人成纤维细胞、淋巴细胞和IL-2。在该模型中，淋巴细胞增殖良好，分泌多种类型的细胞因子，如IFN-g超过10天。(4)初步免疫系统模型的建立：我们重建了一个初步的高级免疫系统模型，该模型由三层组成，即树突状细胞层（下）、癌组织层（中）和淋巴组织层（上）。在这个模型中，我们可以在显微镜下观察到树突状细胞向癌组织的迁移，并通过树突状细胞的迁移实时捕获坏死的癌细胞。我们还通过测量树突状细胞分泌的IL-12浓度来简单、快速地评估树突状细胞的免疫质量。我们成功地利用从手术切除的标本中获得的各种类型的细胞来开发几种类型的人工组织。然而，在这个研究期间，我们并没有成功地将这些人造组织与人造血管连接起来。少

This study was focused on the construction of "a higher in vitro immune system model" by combining several types of artificial tissues, including connective tissues, carcinoma tissues, lymphoid tissues, and blood vessel. To perform the research plan, a new research area "tumor technology" was organized by collaboration of "Division of tumor immunology" and "Division of medical technology".Data obtained for duration of this study are as follows:(1) Development of artificial connective tissue: we reconstituted an in vitro connective tissue model which is made up of human fifroblasts and human umbrical vein-derived endothelial cells (HUVEC) in a type-I collagen gel. In this model, HUVEC migrate and form nets of fine blood vessels without addition of any special growth factors.(2) Development of artificial carcinoma tissue: we reconstituted two types of artificial carcinoma tissue. One is a soft tissue type in which we are able to observe microscopically the growth of the two cell types (c … More arcinoma cells and fibroblasts) in real time for more than 30 days. Another is a hard tissue type which is prepared for immunohistochemical analysis. In this model, we can control the growth pattern of carcinoma cells, i.e., expansive or invasive growth, by combination of cytokines used.(3) Development of artificial lymphoid tissue: artificial lymphoid tissue is organized in a type I collagen net supplemented with OKT-3 and fibrinogen, human fibroblasts, lymphocytes, and IL-2. In this model, lymphocytes well proliferate and secrete several types of cytokines such as IFN-g for more than 10 days.(4) Development of preliminary immune system model: we reconstituted a preliminary higher immune system model which is organized in three layers, i.e., dendritic cell layer (lower), carcinoma tissue layer (middle), and lymphoid tissue layer (upper). In this model, we are able to observe microscopically migration of dendritic cells into carcinoma tissue and capture of necrotic carcinoma cells by migrating dendritic cells in real time. We also estimate simply and quickly immunological quality of dendritic cells by measuring IL-12 concentration secreted by dendritic cells.We succeed in developing several types of artificial tissues using various types of cells obtained from surgically resected specimens. However, we did not succeed in connecting these artificial tissues with artificial blood vessel within this research period. Less