Use for research materials in Pathology--application for molecular pathology

病理科有很多机会从外科手术和尸体解剖中获取材料。然而，它们的用途可能仅限于组织学诊断和其他用于其他目的的试验是必要的。本实验是对l.fresh, 2。3.用于病理研究的固定病理材料。在1。人类胎盘的培养上清含有人类鳞状细胞系的生长因子，并进一步研究了哪些因素可能是刺激这种生长的必要因素。对于肺小细胞癌，l -半胱氨酸、转铁蛋白和二硫根碱也具有生长活性，其中l -半胱氨酸最重要。在2。对Rupp和Locker的方法进行了改进，可以有效地从退行性组织中大量提取RNA。在3。初步探讨了福尔马林固定组织退化的机制。提取和纯化的DNA在10%福尔马林中固定时没有受到损伤。由于脉冲场凝胶电泳和组蛋白-DNA重组实验显示，采用常规DNA提取方法提取福尔马林固定组织时，对组织的损伤程度相同，因此机械损伤不如降解机制重要。低温固定和EDTA的加入抑制了DNA的降解。在福尔马林固定期间，酶活性如DNA酶可能对DNA降解负责。以福尔马林固定组织和石蜡包埋组织作为小鼠免疫原，制备了针对生殖细胞和生殖细胞肿瘤的单克隆抗体。几个试验可能会使用许多病理标本进行病理研究。然而，为了多用途使用它们，可能需要进一步的实验。

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.