| Research Abstract |
Department of Pathology has many chance to get materials from surgical operation and autopsy. However, their uses may be limited to histological diagnosis and other trials to use for another purpose must be necessary. Present experiment was undertaken to examine how to use l.fresh, 2.unfixed, and 3.fixed pathological materials for pathological research. In 1., the culture supernatant of human placenta contained the growth factor for human squamous cell lines and it further was examined which factor might be essential for that stimulation. For lung small cell carcinomas, L-cysteine, transferrin, and bathocuproine disulfo hate had also growth activity and L-cysteine was most important. In 2., extraction method for RNA was examined and modification of Rupp and Locker's method might be effective for massive extraction from degenerated tissues. In 3., the mechanism of degeneration of formalin-fixed tissues was examined basically. The extracted and purified DNA was not damaged when it was fixed in 10% formalin.
Because pulsed field gel electrophoresis and histone-DNA recombination experiment revealed the same damage to formalin-fixed tissues when it was extracted by usual DNA extraction method, the mechanical damage was not so important as the mechanism of degradation. Fixation in low temperature and addition of EDTA inhibited the degradation of DNA. Enzymatic activity such as DNase during formalin fixation might have responsibility for DNA degradation. Monoclonal antibodies which were specific to germ cells and germ cell tumors, were generated from formalin-fixed and parrafin-embedded tissues as mouse immunogen.
Several trials might become possible to use many pathological specimens for pathological research. However further experiments may be necessary for multi- purpose use of them.
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