Description: DOS16 is an adherent human cell line derived from retro-peritoneal soft-tissue sarcoma (leiomyosarcoma with high grade spindle histology) from a 37-year-old female patient with Li Fraumeni Syndrome. 

DOS16 is positive for alterantive lengthening of telomeres (ALT+).

Description Key Words: Human sarcoma 

Also Known As: No other names

Organism: Human (Homo sapiens) 

Strain: N/A

Tissue: Metastatic site: Retroperitoneal space

Growth Properties: Adherent doubling time 65-72 hours.

Morphology: Cells appear spindle shaped to polygonal with short cytoplasmic processes.

Growth Medium: RPMI 1640 (with 2mM L-Glutamine + 25mM HEPES) + 20% Foetal Calf Serum 

Resuscitation: Remove protective cryoflex layer around the ampoule prior to thawing. Thaw the ampoule by gently agitating in a 37°C waterbath; thawing should be rapid (around 2 minutes). A centrifugation step to remove the cryoprotectant after thawing is necessary for this cell line. Recovery from thaw may take 3 days.

Subculturing Procedure:

Medium Renewal: 2-3 times per week. 

Subcultivation Ratio: 1:2 – 1:4 Seeding density 1.3 x10⁴cells/cm². Split subconfluent cultures (70-80%). Harvest the cells using 0.05% Trypsin/EDTA at 37°C for 5 minutes.

Culture Conditions: Incubate the culture at 37°C with 5% CO₂.

Cryoprotectant Medium: 10% DMSO + 90% FCS

Handling Procedure for Frozen Cells: Upon receipt, frozen ampoules should be transferred directly to liquid nitrogen storage without delay, if not to be used immediately. Storage at -80°C may result in loss of viability.

Additional Information:

• Cell line positive for alternative lenghtening of telomeres (ALT+) (PubMed 19935656)
• Near triploid karyotype
• Derived from patient with Li-Fraumeni syndrome
• TP53 mutation at nucleotide 637 C>T, i.e. Exon 6, codon 213

Depositor: Professor Philip Crowe, University of NSW – Australia

References:

Das Gupta P (2007) Gene profiling in soft tissue sarcoma: predictive value of EGFR in sarcoma tumour progression and survival. PhD thesis, UNSW, Sydney. https://doi.org/10.26190/unsworks/20387

Henson, J., Cao, Y., Huschtscha, L. et al. DNA C-circles are specific and quantifiable markers of alternative-lengthening-of-telomeres activity. Nat Biotechnol 27, 1181–1185 (2009). https://doi.org/10.1038/nbt.1587

Dagg RA, Pickett HA, Neumann AA, Napier CE, Henson JD, Teber ET, Arthur JW, Reynolds CP, Murray J, Haber M, Sobinoff AP, Lau LMS, Reddel RR. Extensive Proliferation of Human Cancer Cells with Ever-Shorter Telomeres. Cell Rep. 2017 Jun 20;19(12):2544-2556. doi:10.1016/j.celrep.2017.05.087. PMID: 28636942. 

Cellosaurus: CVCL_C1HQ