A "NEGATIVE" FACTOR FOR GLIOMA CELLS FROM LEPTOMENINGEAL AND MESENCHYMAL CELLS

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The research project was aimed to find out a "suppressor agent" for human gliom cell growth from tissue culture medium of leptomeningeal and/or other cells.Explant cultures of human skin, leptomeninges, meningiomas and gliomas, and of newborn rat leptomeninges were newly performed for the present project. All explant cultures except human leptomeninges were successfully subcultured and able to examine their conditioned media. The basic target cells of the conditioned media for growth assay was a permanent human glioma cell line KNS81 and the growth assay was performed by a colorimetric assay using MTT.Any inhibiting activy was not detected in the conditioned media even in the mixed cultures of the explant cultures and KNS81, although the medium from skin fibroblast was less potential for cell growth than the others. The cells were rinsed two times with a serum-free medium prior to preparing conditioned medium, yet it was insufficient for complete removal of serum factors used as their growth media. It was mandatory to establish serum-independent cell lines for studying growth inhibitors.A human glioma cell line growing in a serum-free medium was successfully isolated from a permanent cell line, KNS 81 and resistered as KNS 81-FD.The autocrine growth factors of KNS 81-FD were intracellularly researved and were massively released extracellularly trypsinisation.The cell growth and morphology were depended on the extracellular matrix. Among of human extracellular matrix proteins, plasma fibronectin and laminin enhanced cell growth, while tenascin suppresses cell growth and spreading, and collagen 1 was intermediate. The results indicated that extracellular matrix proteins act as a key role for cell growth and morphology.