Ht 29 colon cancer cell line

Anti-proliferative activity of the synthesised 3-phenylthiazolo[3,2-a]benzimidazoles 4a–d was evaluated at Stem Cell Therapy and Tissue Reengineering Program, King Faisal Specialized Hospital and Research Center, Riyadh, Saudi Arabia. In vitro anti-proliferative activity was measured by the cell growth inhibition assay. This assay was conducted by use WST-1(water soluble tetrazolium-1) reagent²⁶ for determination of IC₅₀ for each compound. HT-29 colon cancer cell line and MDA-MB-468 triple negative breast cancer cell line were purchased from the American Type Culture Collection. Cells were maintained in RPMI 1640 (Sigma-Aldrich, St. Louis, MO), supplemented with 10% FBS (Fetal Bovine Serum) (Lonza Group, Basel, Switzerland), 100 IU/mL penicillin, 100 mg/mL streptomycin, and 2 mmol/L L-glutamine (Sigma). Cells were seeded into 96-well plates at 0.4 * 10⁴/well and incubated overnight. The medium was replaced with fresh one containing the desired concentrations of the test compounds. After 48 h, 10 μl of the WST-1 reagent were added to each well and the plates were re-incubated for 4 h at 37 °C. The amount of formazan was quantified using ELISA (Enzyme Linked Immunosorbent Assay) reader at 450 nm.

The human HT-29 colon cancer cell line was obtained from the American Type Culture Collection (Manassas, VA, USA). The cells were maintained in Dulbecco's modified Eagle's medium (4.5 g/l glucose), supplemented with 10% fetal calf serum, l-glutamine and antibiotics (Biological Industries, Beit Haemek, Israel) at 37°C with 5% CO₂ in a humidified incubator.