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|
|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.