It is very important that 3T3-L1 cells should be initiated at the recommended seeding density of only 3 X 103 cells/cm2 and should be sub-cultured before the culture reaches 70% to 80% confluence, or when the cells reach 5 X104 to 6 X104 viable cells/cm2. The low density of these cultures will make the culture flasks appear sparse, this is normal. The lot specific information on the Certificate of Analysis will provide the number of cells/vial and expected viability to determine the correct growth area to use. Viability of CL-173 may not be determined by solely by attachment because floating cells during recovery from cryopreservation are viable and normal. The floating cells should be retained by gentle centrifugation and added back to the adherent population.
The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, ATCC® 30-2002 ™, supplemented with bovine calf serum to a final concentration of 10%. It is very important when recovering and subculturing the 3T3- L1 cells that the CO2 environment is properly adjusted for the sodium bicarbonate concentration in the medium. DMEM formulations other than ATCC’s formulation may contain higher sodium bicarbonate (3.7 g/L) and require an increased CO2 level (7-10%) when culturing these cells.
|Date Created||02/21/2014 07:28 AM
|Date Updated||02/21/2014 08:04 AM