Clonal Growth of Hum an Acute Leukem ia Cells in Serum -free Methylcellulose Medium

摘要

The clonal growth of human acute leukemia cell line (K562) and acute myeloid leukemia cells in the serum free culture (SFC) was studied in order to establish a SFC system which could replace the effects of serum by using semi solid methylcellulose culture technique. Our results showed that the clonal growth of K562 cells in semi solid culture was dependent on exogenous serum. The K562 could be grown in SFC supplemented with 4 major replacing substances. The multifactor and multilevel orthogonal experiment demonstrated that the colony formation was statistically influenced by the 4 replacing substances at various concentrations ( P <0.01). Among them, bovine serum albumin had greatest effect on clonal growth of K562 cells with the optimal concentration being 15 mg/L, followed by transferring, cholesterol and insulin with their optimal concentrations being of 150 mg/L, 7.8 mg/L and 7.0 mg/L respectively. SFC system was formed with the 4 substances at their optimal concentrations. Colony formation of the blast cells in 10 patients with acute myeloid leukemia was observed in this SFC system. There was a heterogeneity of acute myeloid leukemia cells among the 10 patients in response to the growth substances. In SFC system, there was a linear relationship between the number of the clonal formation and the count of the added cells, indicating the colony growth of the cells. Primary acute leukemia cells maintained in SFC system in 10 cases could completely form clones. The colony formation number in some cases in SFC system was more than that of the serum containing culture. The SFC system could partially replace the serum for study of the clonal formation of human leukemia cells.