Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease
liposarcoma; well-differentiated
Age
68 years
Gender
female
Ethnicity
Caucasian
Applications
The MDM2 amplification present in this cell line can be used as a model for testing new drugs. This cell line is one of the rare liposarcoma cell lines available.
Storage Conditions
liquid nitrogen vapor phase
Karyotype
49~53,xx,+3;7r [cp8]/49~52,XX,-7,-22,+4~7r, +mar[6]/q8~106, idemx2[cp11].ish r mar (1;4;10;amp12q14-15;15) (wcpl+; wcp 4+; wcp 10+, wcp 12+, amp MDM2, amp CDK4; wcp 15+). rev ish amp (1q24’ 1q31, 4p16, 10p15, 12q14, 12q21, 15q26)
Images
Derivation
The 93T449 cell line was established from a well-differentiated liposarcoma from the retroperitoneum of a 68 year old female.
Karyotypic and molecular characterization of this liposarcoma demonstrated the presence of ring and large marker chromosomes containing MDM2, CDK4 and HMGA2 amplification.
Complete Growth Medium
The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, ATCC 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (ATCC 30-2020) to a final concentration of 10%.
Subculturing
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Remove and discard culture medium.
Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37.0°C to facilitate dispersal.
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
Transfer cell suspension to a centrifuge tube and spin at approximately 125 xg for 5 to 10 minutes. Discard supernatant.
Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
Incubate cultures at 37.0°C.
Subcultivation ratio: A subcultivation ratio of 1:3 to 1:6 is recommended. Medium renewal: every 2 to 3 days
