||Suspension (Some Adherent Cells)
||This myelomonocytic leukemia macrophage-like cell line was derived from a BALB/c mouse
||The cells exhibit only weak effector activity in antibody dependent cell mediated cytotoxicity.
||Lysozyme; granulocyte colony stimulating factor; G-CSF; interleukin-3; IL-3
||It produces the contitutive enzyme lysozyme, interleukin-3 and the granulocyte colony-stimulating activity (CSA). Its growth is inhibited by concentrations of LPS as low as 4.0 ng/mL and blocked completely at higher concentrations. Dextran sulfate also inhibits growth in concentrations of 30 to 40 ug/mL. Production of lysozyme and CSA is not inhibited or is actually enhanced during inhibition of cell growth. The cell surface bears receptor for immunoglobulin and complement. WEHI-3 lines exhibit only weak effector activity against sheep erythrocytes or the tumor target EL-4 in an antibody-dependent cell mediated cytotoxic system.
||Iscove's Modified Dulbecco's Medium (IMDM) contains 4 mM L-glutamine, 4500 mg/L glucose, and 1500 mg/L sodium bicarbonate + 0.05 mM 2-mercaptoethanol + fetal bovine serum to a final concentration of 10%.
||Cultures can be maintained by the addition of fresh medium or replacement of medium.
Alternatively, cultures can be established by centrifugation with subsequent resuspension at 2 X 10e5 viable cells/mL.
Maintain cell density between 2 X 10e5 and 2 X 10e6 viable cells/mL. Adherent cells may be harvested by scraping.
||Atmosphere: air, 95%; carbon dioxide (CO2), 5%
||95% FBS + 5% DMSO (Dimethyl sulfoxide)
|Thawing Frozen Cells:
||SAFETY PRECAUTION: Is highly recommend that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that
some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vessel exploding or blowing off its cap with dangerous force creating flying debris.
1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
3. For cells that are sensitive to DMSO is recommended that the cryoprotective agent be removed immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 7 minutes.
4.Discard the supernatant and Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio).
5. Incubate the culture in a appropriate atmosphere and temperature (see "Culture Conditions" for this cell line).
NOTE: It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
||Cancer Res. 37: 546-550, 1977;
Differentiation 11; 59-63, 1978;
Blood 59: 761-767, 1982
||J.M. Heard, Laboratoire d'Immunologie et virologie de Tumeurs, hospital Cochin, Paris, through Dr. Christine Blaineau, Instituto de Biofisica, UFRJ, Rio de Janeiro.