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Authorized Uses of Cell Biologics’ Products
C57BL/6 Mouse Bone Marrow Macrophages from Cell Biologics are distributed for research purposes only. Our products are not authorized for human use, for in vitro diagnostic or therapeutic procedures. Transfer or resale of any Cell Biologics’ cells or products from the purchaser to other markets, organizations or individuals is prohibited by Cell Biologics without the company’s written consent. Cell Biologics’ Terms and Conditions must be accepted before submitting an order.
Disclaimer Investigators should handle the cells with caution and treat all animal cells as potential pathogens, since no test procedure can completely guarantee the absence of infectious agents.
Cell Biologics’
Primary Macrophage Culture Protocol
All cell culture procedures must be conducted in a bio-safety cabinet.
Any and all media, supplements, and reagents must be sterilized by filtration through a 0.2 µm filter.
Use aseptic technique to prevent microbial contamination.
Medium Review the information provided on the Cell Biologics website about appropriate culture media (e.g. serum and other supplements).
Use pre-warmed (37°C) cell culture media (30-50 ML, Catalog No. M3368) to seed cells and when changing media.
Coating of Cell Culture Plates or Dishes Coat sterile culture dishes or flasks with Gelatin Coating Solution (Catalog No. 6950, Cell Biologics) for 2 min, then aspirate the excess solution.
Cell Recovery from Cryovial
Thaw the cells quickly (<2 min) in a 37°C water bath until just prior to complete thawing.
Wipe the outside of the vial with 70% ethanol.
Use 1 ml Pipette Tip to resuspend cells and transfer cells to a 15 ml sterile tube.
Add 5 ml of pre-warmed Cell Biologics’ Cell Culture Medium (Catalog number M3368, Cell Biologics) gently to the side of the tube and slowly pipette up and down 2 to 3 times to resuspend the cells.
Centrifuge the cells at 200 x g at 4°C for 5 min.
Carefully remove the supernatant and resuspend in 7 mL pre-warmed Maintenance Medium (Catalog No. M3368, Cell Biologics).
Carefully pour cell suspension into the cell culture plate or dish.
Recommended Cell Seeding: 0.7-0.8 million cells are seeded per well of a 12-well plate or 1-1.5 million macroPlace a plate in a humidified, 5%-CO2 incubator at 37°C until experiments.
Change fresh cell culture medium every 24-48 hours. Cells should be checked daily under a microscope to verify appropriate cell morphology.
Proceed with experiment assays.
Note: Please send us the cell images (>90% confluence) if you have any question or problem with cultured cells.
Publication:
mBio. 2021 May-Jun; 12(3): e01227-21.
mSphere. 2022 Jan-Feb; 7(1): e00934-21.
