日本血吸虫培养细胞磷酸酶细胞化学的动态变化

Cytochemical Studies on Dynamic Changes of Phosphatase in the Cultured Cells from Schistosoma Japonicum

目的 :研究体外培养的日本血吸虫成虫细胞碱性磷酸酶 (AKP)和酸性磷酸酶 (ACP)活性的变化规律。方法 :将虫龄 2 6d的日本血吸虫成虫细胞接种于小盖玻片上 ,培养于RPMI16 4 0含 2 0 %小牛血清附加常量抗生素的培养基中 ,培养 7,14 ,2 1,35和 4 9d时 ,分别运用改良的Gomori钙 -钴法和Gomori硫化铅法进行AKP和ACP染色。结果 :随着培养时间延长 ,日本血吸虫成虫培养细胞的AKP和ACP活性均渐减弱。培养 2 1d后 ,所有培养细胞的AKP活性开始减弱 ;培养至 4 9d ,AKP阴性反应细胞所占比例增大 ,但仍有较多细胞具AKP活性 ;而ACP活性在培养 14d时 ,有的细胞显著增强 ,有的却明显减弱 ;培养至 35d ,所有细胞的ACP活性减弱 ,并出现ACP阴性反应的空泡状细胞 ;至 4 9d ,大部分细胞的ACP反应为阴性。结论 :随着培养时间延长 ,日本血吸虫培养细胞的AKP、ACP活性均减弱 ,AKP、ACP活性的强弱可以作为评价血吸虫细胞培养条件优劣的指标 ;培养细胞呈分批、逐步退化。

Objective: To study the changing regularity of activities of alkaline phosphatase (AKP) and acidic phosphatase (ACP) in the cultured cells from adult Schistosoma japonicum(S.japonicum). Methods: The cells obtained from 26 d adult S.japonicum were inoculated on the small glass coverslips. After cultured for 7, 14, 21, 35, and 49 d in RPMI1640 containing 20% calf serum and a moderate amount of antibiotics, the cells were stained to show the activities of AKP and ACP by using Gomori-Ca-Co and Gomori-lead sulfide methods.Results: With the prolongation of cultural time, the activities of AKP and ACP in the cells were got weaker gradually. The activity of AKP in all the cells was decreased at 21 d. Although the proportion of AKP-negative cells was increased, but a few cells possess the activities of AKP when cultured for 49 d. The activity of ACP obviously increased in some cells or decreased in the other cells when cultured for 14 d, and the activity was decreased in all cultured cells and there were some negative cells were as vacuole when cultured for 35 d. The ACP reaction was negative in majority of the cells cultured for 49 d. Conclusion: The activities of AKP and ACP in the cultured cells from adults declined gradually along with the prolongation of the culture time. The levels of AKP and ACP activities could be used to estimate whether the culture conditions were appropriate. The cultured cells from S. japonicum degenerated progressively and group by group.