||Mouse; Dba Subline 212
||8 Month /
||established from the methylcholanthrene-induced tumor in a DBA strain mouse (subline 212, 8-month-old female) in 1949; the cells were propagated by heterotransplantation into DBA/2 mice; an in-vitro suspension culture was first reported in 1966
||cells are used for cytotoxicity studies
||Yes, in nude mice
||Dulbecco's modified Eagle's medium with 2 mM L-glutamine, 4.5 g/L glucose and 10% of horse serum.
||Cultures can be maintained by the addition of fresh medium or replacement of medium.
Alternatively, cultures can be established by centrifugation with subsequent resuspension.
Start cultures at 5 x 10 4 viable cells/mL.
Population Doubling Time: 8 to 10 hrs
|Subculturing Medium Renewal
||Add fresh medium (20% to 30% by volume) every 2 to 4 days.
||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).
||J Nat Cancer Inst 1949; 10:179; J Nat Cancer Inst 1966;36:405
Law LW, et al. Observations on the effect of a folic-acid antagonist on transplantable lymphoid leukemias in mice. J. Natl. Cancer Inst. 10: 179-192, 1949. PubMed: 18143260
Moore GE, et al. Suspension Cell Culture and In Vivo and In Vitro Chromosome Constitution of Mouse Leukemia L1210. J. Natl. Cancer Inst. 36: 405-421, 1966. PubMed: 18630317
Himmelfarb P, et al. Growth of Colonies of. Murine Leukemia L1210 in Vitro. Cancer Chemother. Rep. 51: 451-3 1967
Young SW, et al. Gadolinium(III) texaphyrin: a tumor selective radiation sensitizer that is detectable by MRI. Proc. Natl. Acad. Sci. USA 93: 6610-6615, 1996. PubMed: 8692865
||Banco de Células do Rio de Janeiro