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本文报道使用高渗氯化钠溶液制备脂肪细胞空泡。得到的空泡用Harris 苏木素染色不着色,说明空泡中基本上不含细胞核。用苏丹Ⅲ染色呈极浅的橙红色,表明空泡中含少量脂质。细胞标志酶活力测定表明:空泡的碱性磷酸酯酶活力比完整细胞的高3.5倍,琥珀酸脱氢酶活力和还原辅酶Ⅱ-细胞色素c 还原酶活力均极低。空泡中DNA 含量几乎测不出。以上结果均表明这样得到的空泡,细胞器的含量很低。经测定,空泡对半夏蛋白及胰岛素的结合能力与完整细胞相同。根据组织化学鉴定、细胞器标志酶及细胞膜受体测定说明,用高渗法制备的脂肪细胞空泡主要成分是细胞膜,且保持了膜上受体的活力,适用于膜受体的结构和功能的研究。
This article reports the use of hypertonic sodium chloride solution to prepare adipocytes vacuolar. The resulting vacuoles were stained with Harris hematoxylin, indicating that the vacuoles were essentially free of nuclei. Very light orange-red staining with Sudan III showed that the vacuoles contained a small amount of lipid. Cell marker enzyme activity assay showed that vacuolar alkaline phosphatase activity 3.5 times higher than intact cells, succinate dehydrogenase activity and reduced coenzyme Ⅱ - cytochrome c reductase activity are extremely low. Void DNA content is almost undetectable. The above results show that the resulting vacuoles, organelles, are low in content. It has been determined that vacuoles have the same binding capacity to the albumin and insulin as intact cells. According to histochemical identification, the organelle-labeled enzyme and cell membrane receptor assay showed that the main component of adipocyte vacuoles prepared by hypertonic method is the cell membrane, and maintained the activity of the receptor on the membrane, which is suitable for the structure and function of the membrane receptor the study.