Objective To isolate L1210 cell lines by the single cell cloning technique and to compare some cytology characteristics of L1210 cells and clonal cells.Methods By scanning electron microscope we observed cell superficial structure; DNA content analysis by flow cytometry; the protein distribution analysis by immunohistochemistry. Results Clonal cells L2B9, L2H7,L2H8,L3B12 and L3E11 had a lot of tiny crests and tiny folds, but difference in density, L2C9 had abundant microvilli and L3F9 had massive prominency. DNA contents were 8.8,8.3,8.3,8.5,8.6,8.7,8.9,9.0pg for L1210 and seven sub-lines L2H7,L3F9, L2H8,L2C9,L3B12,L2B9, L3E11,respectively. Statistic analysis indicated significant difference ( P <0.01).Main range of the chromosome number of L1210 cells were 62-63, chromosome number of the clonal cells were 48,58,64,66,62,63,58 of L2B9, L2C9, L2H7,L3B12,L3E11,L3F9,L2H8 respectiverly. With immunohistochemistry different clonal cells had different reactions to 10 kinds of antibodies, and the reaction positive rates of clonal cells were higher than L1210 cells.Conclusion Seven sub -linesshoweddifferentproperties.