Studies On The Cultured Conditions Of Cells From Schistosoma Japonicum And Variant Studies On The Content Of Antigen In The Cells From Schistosoma Japonicum

The effects of complex media, extracelluar matrix (ECM) and serum concentration at 3 different values on the activities of succinodehydrogenase (SDH), lactatede dehydrogenase (LDH) and glucose-6-phosphate dehydrogenase (G-6-PDH) in the cells from Schistosoma japonicum (21-day old) were studied and valued in the present paper. Orthogonal tests were used by the paper to study the cultured condition of primary cells from S. japonicum. In the cultured conditions that was selected immunocytochemistry was used to study the variant change of content of antigen in the cultured cells from S. japonicum. The results are following: 1. Selecting of cultured conditions of Schistosoma japonicumThe effects of complex media (TC-199, RPMI-1640 and DMEM) , extracelluar matrix (liver matrix from rabbit, lung matrix from rabbit, and collagen from rat tail ) and serum concentration (5%, 10% and 20%) at 3 different values on the activities of succinodehydrogenase (SDH), lactatede dehydrogenase (LDH) and glucose-6-phosphate dehydrogenase (G-6-PDH) in the cells from S. japonicum (21-day old) were studied by orthogonal tests. The cells from S. japonicum were stained to show the activities of SDH after they were cultured for 5 days, LDH, G-6-PDH after they were cultured for 14 days, and LDH after they were cultured for 28 days, respectively. Then they were observed and photographed by using the microscope (Olympus-BH2). The concentration of SDH, LDH and G-6-PDH was measured by using photometer-image analytical instrument and statistically analyzed. The results showed: Among of SDH, LDH and G-6-PDH could be used to estimate whether the cultured condition of cells from S. japonicum is appropriate. During the early time of the culture, the highest activities of SDH, LDH and G-6-PDH in the cultured cells from S. japonicum were noted when the cells were inoculated on the glass coverslips which had smeared with the biomatrixes from the liver of rabbits and cultured in the medium of RPMI-1640 containing 20% bovine serum . The effect of serum concentration on the activities of the above enzymes in the cultured cells from S. japonicum was predominent, and the differences among the three enzymes were significant (P <0.01 or P <0.05 ) . The effectof ECM on the activities of the above enzymes was the second, and there was significant difference between SDH and LDH (P <0.01 or P <0.05) . The effects of ECM on the activities of G-6-PDH in S. japonicum cultured cells were not stable. The effect of complex media was the weakest, and the differences among the three enzymes were not significant (P >0.05) . After they were cultured for 28 days the highest activities of LDH in the cultured cells from S. japonicum were noted when the cells were inoculated on the glass coverslips which had smeared with the bio-matrixes from the liver of rabbits and cultured in the medium DMEM containing 5% bovine serum . The effect of serum concentration on the activities of LDH in the cultured cells from S. japonicum was predominent, and the difference was significant (P <0.05 ). The effect of ECM on the activities of LDH was the second (P <0.05 ) . The effect of complex media was the weakest, and the difference was not significant (P >0.05) . So during the early time of the culture the activities of SDH, LDH and G-6-PDH in the cultured cells from S. japonicum are strongest when the cells are inoculated on the glass coverslips which has smeared with the bio-matrixes from the liver of rabbits and cultured by either medium of the three containing 20% bovine serum. After 28 days the activities of LDH in the cultured cells from S. japonicum are strongest when the cells are inoculated on the glass coverslips which has smeared with the bio-matrixes from the liver of rabbits and cultured by either medium of the three containing 5% bovine serum. 2. The variant study of content of antigen in the cells from S. japonicumThe cells from S. japonicum were inoculated on the small glass coverslips that had been smeared with liver matrix from rabbit or hadn't been smeared before. They are cultured in the medium of RPMI-1640 containing 20% bovine serum . Cultured cells from S. japonicum were stained to show the content of antigen by immunocytochemistry , the change of antigen in which were observed by microscope, and the content of antigen was quantitatively analysed by HPIAS-2000. The results showed: After immunocytochemical staining the edge of endoplasma showed heavy brown, brown, light brown in the cells of the first and third group, the other region of plasma showed light yellow. The content of antigen was reduced gradually; the content of antigen in the cultured cells (cells of guoup 1) that had been smeared with liver matrix was reduced less slowly than the cells that hadn't been smeared with liver matrix (cells of guoup 3); moreover , the content of antigen in the cells of group 1 was higerthan that of group 3 at the same time, and the differences between the two groups at the time of 21days, 28days, and 35days are significant (P <0.01 or P <0.05) ; the differences between content of antigen in the cells of group 1 after being cultured for 28days and 35days are significant P <0.05), the differences between content of antigen in the cells of group 3 after being cultured for 21 days and 28days, 28days and 35days are significant P <0.05). So antigen in the cells from S. japonicum are mostly labled in or surrounding the cytoplasma; liver matrix has good effect on the maintanence of antigen in the cultured cells from S. japonicum.