Culture conditions for RAW cells

Mouse macrophage RAW–264.7 cell culture

The cell line

The RAW 264.7 cell line was established from a tumor induced by the Abelson murine leukemia virus. The cells will pinocytose neutral red and will phagocytose latex beads and zymosan. They are capable of antibody dependent lysis of sheep erythrocytes and tumor cell targets. LPS or PPD treatment for 2 days stimulates lysis of erythrocytes but not tumor cell targets. The cell line has been updated to the biological safety level 2 as the cells express a retrovirus which is also infectious for humans.

Features

  • Organism: Mus musculus (mouse)
  • Strain: BALB/c
  • Gender: male
  • Tissue: Ascites
  • Celltype: macrophage
  • Growth Properties: monolayer
  • Risk group: 2

Culture conditions

  • Medium: D-MEM + 10% FCS + 1 % PenStrep oder RPMI 1640 + 2 mM L-glutamine + 10 % FCS
  • Split 1:5 when 80% confluent

Splitting procedure

  • Pre-warm PBS, medium & 0.05% Trypsin to 37°C
  • Remove medium from the cells and wash 2 x with PBS
  • add 1 ml Trypsin per T–75 flask and incubate for 5 min at 37°C
  • bump flask slightly to shake of the cells; check in the microscope
  • add 10 ml medium per T–75 flask and resuspend extensively to have single cells
  • give 2 ml to 8 ml fresh medium (1:5) for a T–75 flask and grow at 37°C, 5% CO2