The ACHN cell line was initiated in November, 1979 from the malignant pleural effusion of a 22-year-old Caucasian male with widely metastatic renal adenocarcinoma (autopsy confirmed). Cells were seeded directly to culture flasks in Eagle''s MEM with 10% FBS, then maintained and passaged 150 days in flasks. Cells were then inoculated subcutaneously into nude mice. After 4 weeks, palpable, locally invasive tumors were noted. Both the original cells (ACHN) and those recovered from nude mouse tumors were growth-inhibited by human interferons.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37¡ãC to facilitate dispersal.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37¡ãC.