Studies on Human Lymph Node by Clinical Tissue Culture and Fluorochrominized Tissue Culture

Abstract

For the lymph-node tissue culture, most works were carried out by cover-slip method with classic media and culture technics up to this time. Therefore, we have studied human lymph-node by means of clinical tissue culture as well as fluorochrominized tissue culture devised in our laboratory, and also applied these technics to clinic.Method: 1. Clinical tissue culture. After a drop of serum was spread out on a HIRAKI's tissue culture plate (depth, 200μ), the tissue fragment is placed in the center of serum and a drop of V. B12 (100γ/cc) was added. A cover slip was laid on the plate and sealed with paraffin, then, the plate was placed in a incubator.2. Fluorochrominized tissue culture. In the medium composed of a drop of serum and a drop of Ao. -V. B12 solution (Acridine orange 0.2mg/cc, V. B12 80γ/cc) the tissue frament was cultured, and observed under the fluorescence microscope with OSRAM HBO 200 mercury vapor lamp, exciter filter BG 12 and barrier filter OG 5.Result: 1. Clinical tissue culture. The growth zone of non-pathologic lymph-node is densely packed with cells in the central part, but in the intermediate to the peripheral zones less densely. In lymphadenitis simplex, the cell growth was far superior to the non-pathologic one, while in the patients with reticulosarcoma, tuberculosis, HODGKIN's disease and cancer metastasis the cell growth were mostly poor. In acute lymphocytic leukemia, cells in the growth zone were extremely dense and the boundary is distinctly defined.2. Fluorochrominized tissue culture.The growth zone of non-pathologic lymph-node presented diffusely yellowish green color sparsely intermingled with reddish orange fluorescent spots. As to the pathologic lymph-node with poor cell growth, reticulosarcoma presented yellowish green fluorescence but in HODGKIN's disease fluorescence had more reddish hue, and in cancer metastasis reddish orange fluorescence was found in the explant. In acute lymphocytic leukemia, fluorescence was green in the central part of the growth zone and yellowish green at the periphery.Conclusion: Since there were some definite differences in growth pattern and fluorescence of the respective growth zone of lymph-node tissue culture in various diseases, these culture methods are useful for the diagnosis of the disease with enlarged lymph-node.