| The effects of Spermidine(Spd)on the growth and proliferation of cultured cells were reaseached by observing the influence on succinate dehydrogenase(SDH),the distribution and dynamics changes of nucleolarorganizerregions (NORs) of the cultured cells from Schistosoma japoncum(Ag-NORs)and alkaline/acidic phosphatase (AKP/ACP) in the cultured cells. It was expected to lay a good foundation for the establishment of cell line of 5. japonicum. 1.Effects of SPD on SDH of the cultured cells from S. japonicumThe first experiment: To study the effect of Spd on the growth and proliferation of cultured cells from adult Schstosoma japonicum,cells collected were cultured in medium RPMI-1640 containing 20% calf serum and a moderate amount of antibiotics for 4 days.The cells were directly treated by Spd with concentrations of 0, 25,50,75, 80,90,150,200μm/L for 24 hours in medium RPMI-1640 with free serum respectively.Afterwards,they were moved to original medium.Then,the cultured cells were stained for SDH in 8cultured day.The contents of SDH in the cells of each group were measured by the photometer-image analytical instrument(HPIA-2000).The results showed the contents of SDH were strongest in 75μm/L Spd. The second experiment:the cultured cells were treated with 75μm/L Spd for 0,12,24,6,48,60,72h after cultured for 4 days in routine medium.Samely,stained for SDH on 8 day.The result showed that the cultured cells grow well after treated with 75μm/L Spd for 48h. 2 .Effects of Spd on Ag-NORs of the cultured cells from S. japonicumIn the experiment the cells from S. japonicum were inoculated on the small glass coverslipS. The cells were divided into test and control groups after cultured for 3 days in routine medium. Cells in the test group were treated with with Spd of the concentration of 75|μm/L for 48 hours in the medium without serum.Afterwars,the cells were cultured in routine medium.The culture condition for the cells in the control group was the same as thetest group except for no addition of Spd in the media.The cells were stained according to the routine cytochemical methods regularly from Iweek to 5 week. The three parameters of AgNORs: number of granules, percentage of AgNORs area/Nucleus area, average optical density were measured. Compared with control group, contents of AgNORs were rising by using Spd in the culture. The three parameters of AgNORs were all statisticaly different between control group and test group (P<0.01). It was suggested that Spd have some ability to increase the potential of transformation and proliferation in cultured cells. 3. Effects of Spd on AKP and ACP of the cultured cells from S. japonicumIn this experiment,3 groups were made:Spd group,liver matix group,Spd+liver matix group. The contents of AKP and ACP in the cultured cells of each group were measured by using HPIA-2000. The results showed the contents of AKP and ACP in cells of Spd group were strongest in three groups in 14 cultured days(P<0.01). AKP and ACP in Spd+liver matix group were the weakest in three groups(P<0.01);21 days later, AKP and ACP in cells of liver matix group were stronger than those of Spd group and the concentrations of the two enzymes in the cells of Spd+liver matix group were lower than 2 weeks before.The activities of AKP and ACP in Spd group were descended more sharply than those in liver matix group. It was suggested that liver matix was more stable and steady to advance the cultured cells than Spd.group.There was adverse-operation between Spd and liver matix.lt was proved that Spd could increase the metabolism in cultured cells and give an impetus to cell growth and proliferation. |
PDF Full Text Request