Liposarcoma

Collection of liposarcomas for studying mechanisms for maintaining telomeres and development of cell lines was performed utilizing an IRB reviewed protocol at Fox Chase Cancer Center. The LS2 cell line was derived from a pleomorphic liposarcoma; it was placed in culture after mechanical disruption. LS2 is maintained in RPMI 1640 + Glutamax (Invitrogen) supplemented with 20% FBS (Invitrogen), MEM Vitamin Mixture (BioWhittaker), ITES (Lonza), Penicillin (100U/ml) Streptomycin (100 μg/ml) L-Glutamine (0.29 mg/ml) mixture (Invitrogen), 1mM sodium pyruvate (CellGro) and MEM Eagle Non-essential amino acid solution (Lonza) with 5% CO₂. The LiSa-2 cell line, derived from a poorly differentiated, pleomorphic liposarcoma(12 (link)), was provided by Dr. W Chow (City of Hope, Duarte, CA) and is maintained in DMEM (Invitrogen) supplemented with 10% FBS (Invitrogen), 25 mM HEPES pH 7.3, Penicillin (100 U/ml) Streptomycin (100 μg/ml) L -Glutamine (0.29 mg/ml) mixture (Invitrogen) with 5% CO₂. The SW872 cell line was obtained from ATCC and is maintained as suggested by ATCC in the absence of CO₂, andin Leibovitz’s L15 medium (Invitrogen) supplemented with 10% FBS (Invitrogen), 0.29mg/ml L-Glutamine and 0.1 μg/ml Normocin (Invivogen). The HeLa cell line was maintained in DMEM supplemented with 10% FBS (Invitrogen) and Penicillin (100 U/ml) Streptomycin (100 μg/ml) L-Glutamine (0.29 mg/ml) mixture (Invitrogen) with 5% CO₂.
DNA fingerprints were obtained for T27, the LS2 cell line derived from T27, and the LiSa-2 cell line using the AmpFlSTR Identifier PCR Amplification kit (Applied Biosystems, Foster City, CA) as recommended by the manufacturer. The kit amplifies the amelogenin gender-determining marker and 15 tetranucleotide repeat loci in a single PCR amplification. This combination of markers is consistent with worldwide database recommendations for identity testing. Allele calls were made from peak plots by comparing peaks to known fragment sizes using GeneMapper 4.0 (Applied Biosystems).