動脈硬化症モデルとしての組織培養

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  • 神尾 昭紀
    The Department of Pathology, School of Medicine, Fukuoka University
  • 白石 昌之
    The Department of Pathology, School of Medicine, Fukuoka University
  • 舌間 憲士
    The Department of Pathology, School of Medicine, Fukuoka University
  • 竹林 茂夫
    The Department of Pathology, School of Medicine, Fukuoka University

書誌事項

タイトル別名
  • In Vitro Culture System for Atherogenesis

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Smooth muscle cells from freshly excised tunica media of the thoracic aorta of 6 month old swine were maintained in continuous culture for 6 months in a moist atmosphere of 95% air and 5% CO2 at 37°C. The modified Dulbecco-Vogt culture medium containing 10% newborn calf serum and 1% antibiotic-antimycotic solution was changed every third day. Growth characteristics of the cells and the extracellular ground substances in long-term cultures were examined ultrastructurally. Cells began to grow out from the explants in 10 to 14 days, adhered to the culture flask and became confluent in 4 to 6 weeks. The cells cultured for 3 weeks had few mitochondria, small amounts of rough endoplasmic reticulum, and a large complement of free ribosomes. When the cultured cells grew to confluency, phasecontrast microscopy revealed that they formed a characteristic pattern of "hills and valleys" in which the cells, having an interlacing or overlapped arrangement, were bridged by parallel starts of surrounding monolayer cells. Ultrastructurally, their cytoplasm contained more numerous numbers of mitochondira, rough endoplasmic reticulum and Golgi complex than those in cultured cells at earlier stages of growth. The cultured cells of 6 month old still had myofilaments, fusiform densities, pinocytotic vesicles and basement membrane characterized as smooth muscle cells, although they had prominent rough endoplasmic reticulum, sparse myofilaments, myelin figures or other lysosomal particles of varying densities and glycogen granules.<br>After cultured cells became confluent, they scattered to form small-sized atherosclerotic mound-like colonies. These cellular aggregates gradually became larger, measuring up to 200μ or occasionally 2mm in width after 3 months. Ultrastructurally, those cellular aggregates exhibited 1 to 4 superficial layers of cells and beneath the layers of cells there were extensive amounts of thin and thick fibrils, amorphous electron dense materials, cellular debris and degenerative cells. The thin fibrils, measuring approximately 110Å in width and similar appearance to "elastin microfibrils", were present throughout the growth. The thick fibrils somewhat resembled to basement membrane with a width of approximately 300Å were also located thoughout the matrix of cell culture specimens. Those fibrils did not have the periodicity of collagen. Foam cells did not occur in long-term cultures of the aortic smooth muscle cells. The present in vitro culture system dealed secretion of extracellular material and cellular degeneration led to the formation of nodular protrusions which may be convenience for the study of the morphological features of atherosclerotic lesions.

収録刊行物

  • 動脈硬化

    動脈硬化 7 (1), 85-91, 1979

    Japan Atherosclerosis Society

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