Ovarian cancer cell lines OVCAR-3 (HTB-161, ATCC, Manassas, VA, USA), SKOV-3 (HTB-77, ATCC) and OV90 (CRL-11732, ATCC), astrocytoma cell line E98 [67 (link)] and primary skin fibroblasts C5120 [46 (link)] were cultured in a humidified incubator at 37 °C and 5% CO2. SKOV-3 and E98 cells were cultured in DMEM (Gibco) with 10% fetal calf serum (FCS). OVCAR-3 cells were cultured in RPMI (Gibco) with 20% FCS. Following recommendation by ATCC, OV90 cells were cultured in MCDB 105 (Cell Applications) and Medium 199 (Sigma-Aldrich) (1:1, v:v) with 15% FCS. C5120 cells were cultured in Medium 199 with 10% FCS.
Tumor cell spheroids of OV90 and SKOV-3 cells were formed by the hanging drop method as described earlier [68 (link)]. Cells were detached with trypsin/EDTA and resuspended in complete culture medium containing 1.2 mg/mL methylcellulose (Sigma-Aldrich) with 200 U/mL penicillin/streptomycin (Sigma-Aldrich). Then, 15,000 cells were seeded in 30 µL drops hanging from the inverted lid of a 140 × 20.6 mm petri dish (VWR international). Spheroids were used for experiments after 48 or 72 h.
Tumor cell spheroids of OV90 and SKOV-3 cells were formed by the hanging drop method as described earlier [68 (link)]. Cells were detached with trypsin/EDTA and resuspended in complete culture medium containing 1.2 mg/mL methylcellulose (Sigma-Aldrich) with 200 U/mL penicillin/streptomycin (Sigma-Aldrich). Then, 15,000 cells were seeded in 30 µL drops hanging from the inverted lid of a 140 × 20.6 mm petri dish (VWR international). Spheroids were used for experiments after 48 or 72 h.
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