HL60 Cell Line human

Differentiation studies

HL60 cell line has been used to study the effect of inecalcitol on the expression of cluster of differentiation 38 (CD38). It has also been used to study the role of autophagy in the cytotoxicity of cytarabine (an anti-leukemic drug).

Human Caucasian promyelocytic leukaemia

STR-PCR Data: Amelogenin: X
CSF1PO: 13,14
D13S317: 8,11
D16S539: 11
D5S818: 12
D7S820: 11,12
THO1: 8
TPOX: 8,11
vWA: 16

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.

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

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. 100ul, 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.

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