Description: The HL-60 cell line was derived from peripheral blood leukocytes obtained by leukopheresis of a 36-year-old Caucasian female with acute promyelocytic leukemia. It was among the first long-term suspension cultures of human myeloid leukaemic cells to be established. Approximately 10% of HL-60 cells spontaneously differentiate and differentiation can be stimulated by polar planar compounds butyrate, hypoxanthine, phorbol myristic acid (PMA, TPA), dimethylsulfoxide (DMSO, 1% to 1.5%), actinomycin D, and retinoic acid.

Also Known As:

Species: Human

Tissue: Blood

Growth Properties: Suspension

Morphology: Lymphoblast

Growth Medium: RPMI 1640 + 2mM Glutamine + 10-20% Foetal Bovine Serum (FBS).

Subculturing Procedure: When starting from a frozen ampoule, add thawed cells to a conical based centrifuge tube e.g. 15ml tube. Slowly add 4 ml of culture medium to the tube. Take a sample of the cell suspension, e.g. 100µl, to count cells. Centrifuge the cell suspension at low speed i.e. 100–150 x g for a maximum of 5 mintues. Remove medium and resuspend the cell pellet at a density of 3–5 x 100,000 cells/ml in fresh medium containing 20% serum. Incubate flask at 37 C; 5% CO2. Cell growth after resuscitation is slow, it may take up to 10 days for proliferation to be established. Check daily. Once the culture is established the serum concentration can be reduced to 10%. Maintain cultures between 1-9 x 100,000 cells/ml; 5% CO2; 37 C; at low density or they may differentiate. At ECACC it has been found to be difficult to recover cells of acceptable viability after freezing in 10% glycerol/90% FBS. We recommend using 10% DMSO/90% FBS for this purpose, but spinning out the cells at resuscitation as above to remove the DMSO as it can cause cells to differentiate. After 6 weeks in culture cells may differentiate.

Release Conditions: No

Depositor:

Originator: No

References: Collins et al.,” (1977) Continuous growth and differentiation of human myeloid leukaemic cells in suspension culture. Nature 270:347?349.

Hyperlink to ECACC Cell Line Data Sheet: 98070106