We have established a cell line from peritoneal macrophages of mice treated intraperitoneally with N-methyl-N-nitrosourea. The present communication describes the identification of an RNA-dependent DNA-polymerase activity in a particulate fraction from supernatants of the cell culture. This activity is similar to retroviral Reverse Transcriptase (RT) based on its template specificity and ionic preference. The proof of the retrovirus-like nature of RT was obtained by ultracentrifugation of the pelleted proteins secreted in the medium on a sucrose gradient. The main RT activity was obtained in fractions of 1.14-1.16 g/ml densities, which are comparable to those of type C retroviruses. The presented data support constitutive expression of the retrovirus gene in a chemically transformed cell line.

The Ymnu cell line established by us from peritoneal exudate cells of mice treated with methylnitrosourea is of macrophage origin. We have shown that 54% of these cells possess Fc-gamma receptors and can bind opsonized erythrocytes; 30% of these cells express the nonspecific esterase and 65% the Mac-1 antigen, indicating these cells are dedifferentiated. Treatment of the cells with various differentiation inducers led to time-dependent redifferentiation of the cells. The expression of the nonspecific esterase increased to 51.1% (TPA), 42.5% (RA), 63.6% (DMSO), 40.6% (SB). The fraction of Mac-1 positive cells increased to 90.5% (TPA), 80.6% (RA), 84.5% (SB) and decreased to 52.7% (DMSO). The maximal effects of the chemicals on expression of these two parameters were achieved at different times following treatment. While RA and SB were effective after one day, the maximum effect of TPA was seen at day 5.

Here we describe the establishment of a cell line from peritoneal exudate cells for NMRI-mice treated with methylnitroso urea, which we designate Ymnu. Tests for macrophage specific characteristics have shown that 54% of Ymnu cells possess Fc-gamma receptors, 30% are nonspecific esterase positive and 65% possess the macrophage specific antigen Mac-1, indicating their macrophage origin. Although of macrophage origin, the majority of these cells are round, 13 microns in diameter. The cells grow partially in suspension and have doubling times varying from 3 to 1.75 days depending on serum concentration. Cultures of Ymnu cells achieve after 14 days densities 4 times higher than those achieved by NIH-3T3 cells. Ymnu cells have lost the anchorage dependence of growth and grew very well on semi-solid media. In addition, they possess an oncogenic potential and build tumors when injected subcutaneously in nude mice.