TISSUE IMAGING IN CELL BIOLOGY

继续发展成像和培养技术，为三个- 在三维多细胞系统中，我们已经培养出了活的人类块， 低剪切力旋转壁血管中小鼠淋巴组织 生物反应器（RWV）。淋巴组织碎片在RWV中保持活力 至少两周组织中的细胞迁移到 周边媒体 细胞迁移出组织是可逆的： 来自外周介质的标记细胞重新填充组织， 可以通过用共聚焦显微镜成像来跟踪命运。 迁移 通过将组织碎片包埋在 胶原蛋白凝胶。 细胞迁移完全停止时，组织 包埋在琼脂糖凝胶中。 琼脂糖包埋的组织块 活的：它们消耗氧气和葡萄糖，在培养基中产生IgG， 保留其结构的基本元素，包括富含B细胞的 由T细胞包围的生殖中心。 高密度类似于 在凝胶顶部培养的组织块（组织培养物）。 琼脂糖包埋的组织块的共聚焦成像揭示了一个网络 毛囊树突状细胞的数量。琼脂糖包埋 组织块保留更多的细胞，并具有更少的凋亡细胞核， 非嵌入式控件。 琼脂糖包埋的组织块 感染了HIV-1淋巴组织长期培养 与外周血淋巴细胞一起在双室系统中， 结合成像技术，使我们能够模拟高 免疫系统的复杂性水平，并解决基本问题 淋巴细胞的迁移，归巢和动力学。 如果没有动物 各种免疫疾病的模型，包括艾滋病毒感染导致 艾滋病，发达的模型提供了一个独特的系统，研究发病机制 在人类淋巴组织中。

Continuing to develop imaging and culturing techniques for three- dimensional multicellular systems, we have grown live blocks of human and mouse lymphoid tissue in a low shear force rotating wall vessel bioreactor (RWV). Fragments of lymphoid tissue remain viable in the RWV for at least two weeks. Cells from the tissue migrate into the surrounding media. Migration of cells out of the tissue is reversible: labelled cells from the peripheral media re-populate the tissue and their fate can be followed by imaging with confocal microscopy. Migration of cells may be partially prevented by embedding the tissue fragments in collagen gels. Cell migration is completely stopped when the tissues are embedded in agarose gels. Agarose-embedded blocks of tissue remain alive: they consume oxygen and glucose, produce IgG into the media and retain the basic elements of their architecture, including B cell-rich germinal centers surrounded by T cells. Tall densities are similar that of tissue blocks cultured on the top of the gels (histocultures). Confocal imaging of agarose-embedded blocks of tissue reveal a network of follicular-dendritic cells in germinal centers. Agarose-embedded blocks of tissues retain more cells and have less apoptotic nuclei than non-embedded controls. Agarose-embedded blocks of tissues are productively infected with HIV-1. Long-term culturing of lymphoid tissue together with peripheral lymphocytes in a two-compartment system, in combination with the imaging technique, allows us to simulate the high level of complexity of the immune system and to address basic questions of lymphocyte migration, homing and kinetics. In the absence of animal models for various immune disorders, including HIV infection leading to AIDS, the developed model provides a unique system to study pathogenesis in human lymphoid tissue.