Leukemia cells were seeded at 25.000 cells per well in 100 µL of complete growth medium in 96-well plates and incubated with indicated chemotherapeutic agent or vehicle. T-ALL cells were split every 48 h. Briefly, 20 µL of cells were mixed with 80 µL of fresh culture medium, supplemented with vehicle or chemotherapeutic drugs at the indicated doses.
For the treatment of HCT15 cells, 250.000 cells were seeded in 2.5 mL of culture medium supplemented with the indicated drugs in a 12-well or 6-well plate format, respectively. Cell viability was assessed by counting viable cells based on trypan blue vital dye staining (Invitrogen), according to the manufacturer’s instructions.
For the treatment of organoids, Matrigel and basal organoid medium without growth factors were supplemented with vehicle or 100 U/L of asparaginase and split every 48 h.
All asparaginase experiments were performed using pegaspargase (Oncaspar, Shire Pharmaceuticals, Lexington, MA, USA), an FDA-approved PEGylated form of E. coli asparaginase. Homoharringtonine (SML1091) was obtained from Sigma-Aldrich. All drugs and reagents were used at the indicated concentrations.
For the treatment of HCT15 cells, 250.000 cells were seeded in 2.5 mL of culture medium supplemented with the indicated drugs in a 12-well or 6-well plate format, respectively. Cell viability was assessed by counting viable cells based on trypan blue vital dye staining (Invitrogen), according to the manufacturer’s instructions.
For the treatment of organoids, Matrigel and basal organoid medium without growth factors were supplemented with vehicle or 100 U/L of asparaginase and split every 48 h.
All asparaginase experiments were performed using pegaspargase (Oncaspar, Shire Pharmaceuticals, Lexington, MA, USA), an FDA-approved PEGylated form of E. coli asparaginase. Homoharringtonine (SML1091) was obtained from Sigma-Aldrich. All drugs and reagents were used at the indicated concentrations.