日本血吸虫成虫培养细胞糖细胞化学的研究

目的 :研究日本血吸虫成虫培养细胞内的糖类物质。方法 :将 32d虫龄的日本血吸虫成虫制成细胞悬液 ,接种在小盖玻片上 ,置含 2 0 %小牛血清附加常量抗生素的RPMI 16 4 0培养基中培养 ;至培养第 6d ,用高碘酸雪夫 (PAS)法显示培养细胞中的糖类物质 ;阿新蓝 PAS法显示培养细胞中的酸性粘多糖 ;用淀粉酶处理后的PAS染色以鉴定细胞内的糖原。在Olympus BH2 显微镜下观察 ,统计细胞类型 ,拍照 ;HPIAS 2 0 0 0图像分析仪测定含量 ,统计分析不同细胞类型间的差异。结果 :日本血吸虫成虫培养细胞的类型不同 ,所含的糖类成分有所不同 ,有较多培养细胞内既有糖原 ,也有酸性粘多糖 ,以及中性粘多糖、粘蛋白、糖蛋白、糖脂等其它糖类物质 ;大多数细胞几乎不含糖原和酸性粘多糖 ;另有少数细胞仅含糖原和酸性粘多糖 ,以糖原为主 ,极少数细胞以酸性粘多糖为主。结论 :日本血吸虫成虫培养细胞内既含糖原 ,也含酸性粘多糖 ,以及中性粘多糖、粘蛋白、糖蛋白、糖脂等其它糖类物质

Cytochemical Studies on Carbohydrate of Cultured Cells from Adult Schistosoma Japonicum

Objective: To study the concentration and distribution of carbohydrate in the cultured cells from adult Schistosoma japonicum. Methods: The cells from 32 day old adult S. japonicum were inoculated on glass coverslips and cultured in RPMI 1 640 containing 20% calf serum and a moderate amount of antibiotics. The cells cultured for 6 days were stained to show the concentrations of carbohydrate and acid mucopolysaccharide by using Periodic Acid Schiff reaction (PAS) and Alcian Blue PAS, respectively. Glycogen was also indicated by PAS after the cells were digested by amylase. The stained cells were observed, classified and photographed under a microscope (Olympus BH 2). The concentration of carbohydrate was measured by using photometer image analytical instrument (HPIAS 2000) and analyzed statistically. Results: There were different concentration and distribution of carbohydrate in different types of cultured cells from adult S. japonicum. In a few cultured cells, there were not only glycogen and acid mucopolysaccharide, but also neutral mucopolysaccharide, mucoprotein, glycoprotein and glycolipids. Little glycogen and acid mucopolysaccharide were observed in most of cells. Much glycogen and little acid mucopolysaccharide were detacted in a few cells. Acid mucopolysaccharide took a major ratio in very few Cells. Conclusion: Not only glycogen and acid mucopolysaccharide, but also neutral mucopolysaccharide, mucoprotein, glycoprotein and glycolipids were found in cultured cells from adult S. japonicum.