肝基质对日本血吸虫培养细胞生长的影响

Effects of liver matrix on growth of cultured cells from Schistosoma japonicum

目的研究肝基质对日本血吸虫培养细胞形态、增殖的影响。方法联合法将虫龄为21d的日本血吸虫童虫细胞接种于预先铺敷有肝基质(实验组)和未铺敷肝基质(对照组)的小盖玻片上,培养于常规培养基中,用高碘酸雪夫(PAS)法染色培养细胞,Olympus-BH2显微镜下观察并拍照;并将接种第3天的实验组和对照组培养细胞用胶银法进行核仁组织区相关嗜银蛋白(AgNORs)染色,HPIAS-2000图像分析仪测定AgNORs平均光密度值进行图像分析。结果实验组中的培养细胞其细胞质突起和分裂细胞增多,AgNORs的平均光密度值显著高于对照组细胞(P<0.01)。实验组的细胞分裂方式除常见的一分为二方式外,还观察到一分为三的多极分裂方式;而对照组未观察到分裂细胞。结论肝基质可明显影响日本血吸虫培养细胞的形态,显著促进培养细胞的增殖。

Objective To study the effects of the liver matrix （the bio-matrixes from the liver of a rabbit） on the morphology, growth and proliferation of the cultured cells from Schistosoma japonicum. Methods By using a method of combination, the cells from 21-day old S. japonicum were inoculated on the glass coverslips which had smeared with the liver matrix and cultured in RPMI-1640 containing 20% calf serum and a moderate amount of antibiotics. The cultured cells were stained by using Periodic Acid Sehiff reaction （PAS）. The stained cells were observed and photographed under a microscope （Olympus-BH2）. Simultaneously, the cells cultured for 3 days were stained by the silver colloid technique to show silver stained nucleolar organizer regions （AgNORs）. Average optical density of AgNORs in the cultured cells from S. japonicum was measured by using the photometerimage analytical instrument （HPIAS-2000） and analyzed statistically. The control group was not smeared with the liver matrix. Results The cells cultured on the liver matrixes had more cytoplasmic processes and dividing cells under microscopy. Average optical density of AgNORs was higher in the cells cultured in the liver matrix than that in the control group. There was a significant difference among them （P〈0.01）. Apart from the ordinary binary division of cells, the multipolar division such as tripolar division was also observed in the cultures. Conclusion The liver matrix could evidently influence morphology and better promote division of the cultured cells from S. japonicum.