Human iPSC-Derived Macrophages in a 96-Well Plate Format

Catalog. No. Product Name Format Stock Conc. Storage on Arrival Thawing Instructions Storage Once Thawed
ax0601 Plated Human iPSC-Derived Macrophages (Male) 1x 96-well plate 15,000 cells/well N/A 37°C N/A N/A
ax0606 Plated Human iPSC-Derived Macrophages (Female) 1x 96-well plate 15,000 cells/well N/A 37°C N/A N/A
ax0600 Macrophage Maintenance Medium 250 mL 1x Store at 4°C for 2 months N/A N/A

 

Additional Reagents

Product Name Provider Catalog. No.
Recombinant Human M-CSF Gibco, Life Technologies PHC9501

These reagents must be added fresh for each use to each aliquot of medium.

Preparation of Reagents: 

Recombinant Human Macrophage Colony Stimulating Factor (M-CSF)

  • Prepare a 100 μg/mL stock solution of M-CSF by resuspending the 100 μg of lyophilized powder in 1 mL of Dulbecco’s-PBS (1x) (D-PBS without calcium or magnesium) supplemented with 0.05 % human serum albumin.

Macrophage Maintenance Medium

  • l Upon receipt, store Macrophage Maintenance Medium at 4°C protected from light.
  • Macrophage Maintenance Medium requires supplementing with M-CSF before use.
  • Prepare the volume of Macrophage Maintenance Medium required by adding M-CSF (100 ng/mL). Once M-CSFhas been added the medium is stable for 1 week at 4°C.
  • Pre-warm an aliquot of Macrophage Maintenance Medium to 37°C before use
M-CSF 100 μg/mL 100 ng/mL 50 μL

Maintenance of Human iPSC-Derived Macrophages

Axol iPSC-Derived Macrophages are derived from iPSCs that have undergone an intermediate differentiation to monocytes prior to directed terminal differentiation to macrophages. The cells you receive will have undergone 6 days of macrophage differentiation and should therefore be used within 7 days to avoid overactivation of the macrophages, which typically occurs at day 14.

  • Carefully open the plastic package and place the 96-well plate into a class II biosafety cabinet. Making sure to spray the plate with 70% ethanol and wipe it down.
  • Remove the silicone sealing cap from the plate.
  • Check cells under the microscope.
  • Incubate the plate at 37°C, 5% CO2 for 2 hours.
  • After the incubation period gently replace the half the volume of medium with fresh, pre-warmed 37°C, Macrophage Maintenance Medium.
  • Incubate the plate at 37°C, 5% CO2.
  • To maintain a healthy macrophage culture, replace the half the volume of medium with fresh, pre-warmed 37°C, Macrophage Maintenance Medium every 4 days.
  • Conduct preferred assays within 7 days of receiving the cells.