Product Information

Catalog. No. Product Name Format Stock Conc. Storage on Arrival Thawing Instructions Thawing
ax0666 Plated Human iPSC-Derived Microglia (Female 1x 96-well plate 30,000 cells/well N/A 37ºC N/A N/A
ax0660 Microglia Maintenance Medium + Supplement A, B and C 100 mL + 100 μL + 100 μL + 1 mL 1x Store Microglia Maintenance Medium at 4ºC for 2 months,
Store supplements A, B and C at -80ºC
N/A N/A

Important! Axol Cell Culture Media DOES NOT contain antibiotics or antifungal agents. Axol Bioscience does not recommend the use of antimicrobial agents such as penicillin, streptomycin and amphotericin. Antimicrobial agents should not be necessary if proper aseptic technique is adopted. Recommendations

Recommendations

  •  Recommended cell culture medium: Microglia Maintenance Medium supplemented with Supplement A, B and C

Preparation of reagents

Supplement A

  • Store Supplement A at -80 ºC on arrival.
  • After initial thaw, use 50 μL of Supplement A to make up 50 mL of Microglia
  • Maintenance Medium. Aliquot the remaining 50 μL and store at -80 ºC until required. Do not thaw and refreeze more than once.

Supplement B

  • Store Supplement B at -80 o C on arrival.
  • After initial thaw, use 50 μL of Supplement B to make up 50 mL of Microglia
  • Maintenance Medium. Aliquot the remaining 50 μL and store at -80 ºC until required. Do not thaw and refreeze more than once.

Supplement C

  • Store Supplement B at -80 ºC on arrival.
  • After initial thaw, use 500 μL of Supplement C to make up 50 mL of Microglia Maintenance Medium. Aliquot the remaining 500 μL and store at -80 ºC until required. Do not thaw and refreeze more than once.

Microglia Maintenance Medium

  • Upon receipt, aliquot and store Microglia Maintenance Medium at 4 ºC.
  • Microglia Maintenance Medium requires supplementing with Supplement A, B and C before use.
  • Add the Supplements to the Microglia Maintenance Medium according to the table.
  • Once Supplements A, B and C have been added to the Microglia Maintenance Medium it is stable for 1 week at 4ºC.
  • Before use, pre-warm an aliquot of Microglia Maintenance Medium at 37ºC.
Supplement Final Concentration 50 mL Microglia Maintenance Medium
Supplement A 10 ng/mL 50 μL
Supplement B 100 ng/mL 50 μL
Supplement C 1 x 500 μL

Maintenance of Human iPSC-Derived Microglia

Axol’s Human iPSC-Derived Microglia are derived from iPSCs which have undergone an intermediate differentiation to monocytes prior to directed terminal differentiation to microglia. The cells have undergone 7 days of microglia differentiation and should be cultured for a further 7 days (until day 14) before use and then used within 7 days.

The 96-well plate will arrive in a sealed plastic bag and the wells will be capped with a silicone seal. The Human iPSC-Derived Microglia will be shipped on day 7 post-differentiation from Human iPSC-derived Microglia. Depending on region the shipment should arrive on day 8 or day 9.

  • 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 o C, 5% CO2 .
  • On day 9 gently replace 80 μL of medium with 100 μL of fresh, pre-warmed 37ºC,
  • Microglia Maintenance Medium.
  • Incubate the plate at 37ºC, 5% CO2 .
  • To maintain a healthy microglia culture, gently replace 80 μL of medium with 100μL of fresh, pre-warmed 37ºC, Microglia Maintenance Medium every other day.
  • Culture the Human iPSC-Derived Microglia for a minimum of 6 days before conducting preferred assays. Optimally, Microglia should be assayed within 14 days of receiving the cells.