Htb 88

SK-LMS-1 Human leiomyosarcoma cell line was procured from ATCC (HTB-88, ATCC, Manassas, VA, USA). STS117 Human soft-tissue-sarcoma (STS) primary cell line harboring a loss of function mutation TP53 was derived from patients’ primary extremity STS diagnosed as an undifferentiated pleomorphic sarcoma. STS117 cell line was kindly provided by Dr. R. Gladdy (Mount Sinai Hospital, Toronto, ON, Canada) [17 (link)]. SK-LMS-1 was cultured in EMEM (Wisent Inc., St-Bruno, QC, Canada), STS117 was cultured in DMEM F12 (Wisent Inc.), both supplemented with 10% Fetal Bovine Serum (FBS) (Gibco, Thermo Fisher Scientific, Saint-Laurent, QC, Canada) and 1% Penicillin–Streptomycin solution (Wisent Inc.). SK-LMS-1 and STS117 cells were maintained by subculturing at 80% confluency. Briefly, the medium was aspirated, cells were washed with Phosphate Buffer Saline (PBS) (Wisent Inc.) and were trypsinized with 0.025% trypsin EDTA (Wisent Inc.) for 2–3 min at 37 °C. Once cells are detached, supplemented medium was added to stop the enzymatic reaction and the cell suspension was centrifuged for 5 min at 1500 rpm. The cell pellets were resuspended in the appropriate volume to perform seeding in the microfluidic devices.

Two human uLMS cancer cell lines, SK-UT-1B (ATCC HTB-115™) and SK-LMS-1 (ATCC HTB-88™), and one human uterine smooth muscle cell line HUt-SMC (ATCC PCS-460-011™), were purchased from ATCC (Manassas, VA, USA). uLMS cell lines were cultured in Eagle’s Minimum Essential Medium (EMEM) (ATCC) with 10% Fetal Bovine Serum (FBS) and 1% Pen-Strep. HUt-SMC was cultured in Vascular Smooth Muscle Cell Growth Kit (ATCC) as per the manufacturer’s instructions. All cells were maintained in a humidified incubator under standard culture conditions of 21% O₂ and 5% CO₂ at 37 °C.
γ-secretase activity was blocked by DAPT (Sigma-Aldrich, St. Louis, MO, USA) or MK-0752 (MedChemExpress, Monmouth, NJ, USA) dissolved in DMSO. SK-UT-1B and SK-LMS-1 cells were grown to greater than 80% confluence in 60 mm plates or 6-well plates and treated with DAPT or MK-0752. Vehicle control (DMSO) was used at equal volume to treatment.