[Study on RAPD characteristics of five S180 clonal strains and on biological characteristics of passage cells from S180-S2D9].

Objective: To study the random amplified polymorphic DNA (RAPD) characteristics of five S180, clonal cell strains with 23 primers and the biological characteristics of passage cells from S180-S2D9.
Methods: The DNA of 5 clone strains of S180 including S180-S2D9, S180-S2D7, S180-S1F11, S180-S1H10 and S180-S1B11, was amplified with RAPD-PCR using 23 single primers. The PCR products were resolved with electrophoresis on agarose gel. To analyze genomic DNA with RAPD, the cultured cells in vitro were treated with colchicine for 6 hours. Then, the chromosome numbers of 100 cells were examined under the microscope. The KM mice were injected intraperitoneally with 0.2 ml solution of S180-S2D9 cell, and the growth of ascitic fluid and the life span of the mice were observed. The cultured cells were fixed with 750 ml/L ethanol, and DNA analysis was made by Flow Cytometry.
Results: Three of the 23 single primers resulted in diversity between amplified products from different clonal strains. RAPD character of S180-S2D9 was analyzed by the 3 single primers; RAPD bands of the first generation, 25th generation, 50th generation and 75th generation showed no difference; ANOVA showed that the average numbers of chromosomes of four generations were of no significant difference (P>0.05). The ascites of the KM mice subjected to the intraperitoneal injection of first generation and 50th generation S180-S2D9 cells was bloody, and the survival days of mice were 13-23 d and 13-20 d respectively; ANOVA showed there was no significant difference between the first generation and 50th generation (P>0.05). DNA contents assayed by flow cytometry revealed that DNA corresponding content of the cells is individually 0.3890, 0.3542, 0.3575 and 0. 3984. These results imply that the S180-S2D9 passage cells have not been found to vary a lot.
Conclusion: It is adaptable to give quality control to the clonal cell strains with RAPD.