Complete Growth Medium
Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose, 70%; fetal bovine serum, 10%; LADMAC Conditioned Media (produced from the LADMAC cell line (CRL-2420), 20%
- Remove and discard 75% of culture medium.
- Scrape cells with cell scraper.
- Add appropriate aliquots of cell suspension to new culture vessels.
- Place culture vessels in incubators at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended
Medium Renewal: Every 2 to 3 days
LADMAC Conditioned Medium
LADMAC conditioned medium is made from LADMAC cells (ATCC CRL-2420). LADMAC cells secrete the growth factor colony stimulating factor 1 (CSF-1). CSF-1 is capable of supporting the in vitro proliferation of mouse bone marrow macrophages. The Pannell-Milstein roller bottle apparatus may be used to produce high concentrations of CSF-1 Ref
Subculturing Procedure for LADMAC cells
Cultures can be established by centrifugation with subsequent resuspension at 2 X 105 viable cells/mL. Attached cells may be dispersed by tapping the sides of the flask until cells are loose.
Complete Growth Medium for LADMAC cells
The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%
This medium is formulated for use with a 5% CO2 in air atmosphere.
Conditioned medium harvesting:
- Allow LADMAC cells to become confluent.
- After 5 to 7 days, collect supernate, centrifuge at 125 x g for 5 to 10 minutes
- Filter (200 nM filter) and tore aliquots at –20oC
Culture medium 95%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.