Hek293

The human CRC cell lines HT29 and HEK293 were obtained from the Korean Cell Line Bank (KCLB, Seoul, Korea), and KM12SM-luc was provided by The University of Texas MD Anderson Cancer Center. HT29, KM12SM-luc, and HEK293 were maintained in DMEM (Welgene, Gyeongsan, Korea) with 10% fetal bovine serum (RMBIO, Missoula, MT, USA) and a 1% antibiotic–antimycotic solution (Thermo Fisher Scientific, Waltham, MA, USA).

We were purchased the SKOV3 human epithelial ovarian cancer cell line from the Korean Cell Line Bank (KCLB, Seoul, Korea), and the OVCA429 and OVCA433 cell line were provided by the Korea Gynecologic Cancer Bank through the Bio and Medical Technology Development Program of the Ministry of Science, Information and Communication Technology, and Future Planning (MSIP, Seoul, Korea). The HEK293, TOV112D and OVCAR3 cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA), and the A2780 cell line was procured from the European Collection of Cell Cultures (ECACC, London, UK). SKOV3, TOV112D and A2780 cells were cultured in RPMI-1640 medium (Welgene, Seoul, Korea), and HEK293, OVCAR3, OVCA429 and OVCA433 cells were cultured in Dulbecco’s Modified Eagle’s Medium (Welgene, Seoul, Korea). All culture media were supplemented with 10% foetal bovine serum and 1% penicillin/streptomycin, and cell lines were maintained at 37 °C in a humidified atmosphere of 5% CO₂ and 95% air. Culture medium was replaced with fresh medium every 2–3 days, and cells were used between Passages 5 and 10. OVCA429 and OVCA433 cells were used between passages 10 and 20. The subtypes of the epithelial ovarian cancer cell lines are as follows: SKOV3, serous; TOV112D, endometrioid; A2780, non-specified; OVCAR3, carcinoma; OVCA429, serous; and OVCA433, serous [29 (link)].

The cytotoxicity of the hydrogels was evaluated using WST-8 assays using human embryonic kidney 239 cells (HEK-293, Korean Cell Line Bank, Korea). For direct cytotoxicity tests, HEK-293 cells were seeded into 24-well culture plates at a concentration of 3 × 104 cells per well with minimum essential medium (MEM, WELGENE, Gyeongsan-si, Korea) containing 10% fetal bovine serum and 1% penicillin/streptomycin; then, 5 mg of hydrogel sample was added to the wells and plates were incubated at 37 °C in an atmosphere containing 5% CO₂. MEM medium was used as a negative control, and 10% (v/v) dimethylsulfoxide (DMSO) dissolved in the MEM medium was used as a positive control. After a 48-hour incubation, WST-8 assay reagent (QuantiMax, BIOMAX, Seoul, Korea) was added to each well and absorbance was measured at 450 nm. Cell viability was determined using the following formula: Cellviability(%)=AbsorbanceofcellswithhydrogelAbsorbanceofnegativecontrolcells
All assays were repeated in triplicate for each sample.