鼠胚胎成纤维细胞饲养层及其与肝基质联合对日本血吸虫童虫培养细胞LDH、ACP影响的研究

目的以乳酸脱氢酶(LDH)、酸性磷酸酶(ACP)为评价指标,研究鼠胚胎成纤维细胞(MEF)饲养层及其与肝基质联合对日本血吸虫童虫培养细胞代谢的影响。方法实验分MEF与肝基质联合组(后简称联合组)、MEF组、肝基质组和对照组4组。联合组与肝基质组预先铺敷肝基质,MEF组和对照组未铺敷肝基质,将虫龄21d的日本血吸虫童虫细胞分别接种于预先铺敷或未铺敷肝基质的小盖玻片上。肝基质组和对照组细胞培养于RPMI1640含20%小牛血清附加常量抗生素的常规培养基中,联合组与MEF组的细胞培养于有MEF饲养层的常规培养基中。培养第7d,运用酶细胞化学方法对日本血吸虫童虫培养细胞进行LDH、ACP染色,OlympusBX51显微镜下观察并拍照,HIPAS2000图像分析仪测定其含量,并作统计分析。结果4组童虫培养细胞其LDH、ACP活性不同。LDH、ACP着色按联合组、MEF组、肝基质组、对照组依次变浅。定量分析显示,培养细胞的LDH含量,各组之间两两比较差异具显著性(P<0.01);联合组、MEF组、肝基质组培养细胞的ACP含量两两比较存在显著差异(p<0.01),联合组、MEF组与对照组之间比较差异具统计学意义(P<0.01),而肝基质组与对照组之间差异无显著性(P>0.05)。结论MEF饲养层能促进日本血吸虫童虫培养细胞的代谢,并能与肝基质协同促进培养细胞代谢。

Effect of mouse embryonic fibroblast feeder layer and liver matrix of rabbit on the activities of LDH and ACP in cultured cells from schistosomula Schistosoma japonicum

To study the effect of feeder layer with mouse embryonic fibroblast (MEF) or/and with liver matrix of rabbit on the activities of LDH, ACP in the cultured cells from schistosomula Schistosoma japonicum, the cells from 21 day-old schistosomula were inoculated on small glass coverslips with or without liver matrix smeared previously. Cells with matrix were divided into two groups respectively: one of which was cultured in routine medium that RPMI-1640 containing 20 % calf serum with a moderate amount of antibiotics, the other was cultured in routine medium containing MEF feeder layer. Cells inoculated without liver matrix were also divided into two groups which were cultured in routine medium and MEF co-culture system respectively. So there were four groups in our experiments: liver matrix group, co-culture group, control group and MEF group. The cells were stained to show the activities of LDH and ACP after cultured for 7 days. It was found that the activities of LDH and ACP in the cultured cells were strongest in the cells of co-culture group, then in the MEF group and the liver matrix group, the activities were weakest in the control group. Quantitative analysis showed that activity of LDH had significant difference (P<0.01) among those four groups. The activity of ACP was higher in the co-culture group and the MEF group than the control group (P<0.01); there was no significant difference between the liver matrix group and the control group (P>0.05); but the co-culture group , the MEF group and the liver matrix group were statistically different (P<0.01). It was suggested that the MEF feeder layer can promote metabolism of cultured cells from schistosomula. Further more, the MEF and the liver matrix have a co-effect on the activity of cells from schistosomula.