HL60 cells (ATCC® CCL-240™) were grown in RPMI 1640 medium (Gibco), with 20% FBS + 5% Penicillin-Streptomycin) at 37°C under 5% CO2, supplemented with 10% fetal bovine serum (Sigma) and 1% Penicillin/Streptomycin (HyClone). Cells were maintained below a density of 106 cell/mL. On the first day of the differentiation experiment, cells were seeded at 200,000 cell/mL in 12 well plates (unless stated otherwise) and treated with 1 μM ATRA (all-trans-retinoic acid, Cat#R2625-100MG) to differentiate into either the neutrophil-like cells. Cell differentiation status was confirmed by flow cytometry analysis of CD14 (Biolegend, Cat#367117) and CD11b (Biolegend, Cat#301309).
For the human hematopoiesis dataset, we cultured primary human CD34+ hematopoietic stem and progenitor cells obtained from the Fred Hutchinson Cancer Research Center. Cells were thawed and cultured in StemSpan SFEM II human hematopoietic stem cell expansion media (StemCell Technologies, Cat#02690) supplemented with StemSpan CC100 (StemCell Technologies, Cat#02690) and 50 ng/ml TPO (PeproTech, Cat#300-18-100UG). Cells were allowed to differentiate over the course of 1 week.
For the human hematopoiesis dataset, we cultured primary human CD34+ hematopoietic stem and progenitor cells obtained from the Fred Hutchinson Cancer Research Center. Cells were thawed and cultured in StemSpan SFEM II human hematopoietic stem cell expansion media (StemCell Technologies, Cat#02690) supplemented with StemSpan CC100 (StemCell Technologies, Cat#02690) and 50 ng/ml TPO (PeproTech, Cat#300-18-100UG). Cells were allowed to differentiate over the course of 1 week.