Ovsaho

EOC cell lines MDAH2774, SKOV3, OVCAR3, OVSAHO, OVTOKO and OVISE cells were purchased from ATCC (Manassas, VA). Following tests of these cell lines for immunological markers and cytogenetics, they were also fingerprinted and species was confirmed by IEF of AST, MDH and NP. The cell lines were cultured in RPMI 1640 supplemented with 10% (v/v) fetal bovine serum (ATCC), 100 units/mL penicillin, and 100 units/mL streptomycin (SIGMA) at 37°C in humidified atmosphere containing 5% CO₂. All experiments were performed in RPMI 1640 (ATCC) containing 5% serum.

Clinical specimens of HGSC and normal tissues were collected from the gynecological oncology service of Tzu Chi General Hospital, Hualien, Taiwan, under the approval of the Institutional Review Board of this hospital. Informed consent was given by each subject. The clinical specimens included 9 HGSC specimens (6 derived from the ovary and 3 from the fallopian tube as the primary site), 6 normal ovarian tissues, 5 normal fallopian tube tissues, and 9 normal fimbriae tissues. Normal fimbrial epithelium was carefully removed using a cytobrush and collected in phosphate buffered saline (PBS). Ovarian cancer cells, OVSAHO, SKOV3, and A1847 were purchased from ATCC and cultured in RPMI1640 and Dulbecco’s modified Eagle’s medium, respectively, supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin (PS). The matrigel invasion subclones SKOV3-I6 (six selections for matrigel invasiveness) and A1847-I4 (four selections) were a generous gift from Prof. Lu-Hai Wang of the National Health Research Institutes of Taiwan [32 (link)]. Kuramochi cells were purchased from the Japanese Collection of Research Bioresources (JCRB0098) and cultured in RPMI1640 supplemented with 10% FBS and 1% PS.

The human ovarian cancer cell lines SKOV3 and OVSAHO were purchased from ATCC^® (Manassas, VA, USA) and JCRB Cell Bank (Japan), respectively. Cells were cultured in High Glucose Dulbecco's Modified Eagle's Medium (DMEM) with sodium pyruvate (Euroclone #ECB7501L) supplemented with 10% FBS South America origin EU Approved (Euroclone #ECS5000L), 2 mM L-Glutamine (Euroclone #ECB3000D), 1% penicillin/streptomycin (Euroclone #ECB3001D) and 50 µg/mL uridine (Sigma–Aldrich #U3003) and maintained at 37 °C in a humidified atmosphere with 5% CO₂. For high- or low-glucose experiments, cells were grown for 24 hours in DMEM (Gibco #11966025) supplemented with 10% FBS, 2 mM L-glutamine, 1% penicillin/streptomycin, 50 µg/mL uridine, 1 mM sodium pyruvate (Sigma–Aldrich #P2256) and 25 mM or 5 mM D-(+)-glucose (Sigma Aldrich #G7021). Where indicated, cells were treated with 1 µM N4-[2-(4-Phenoxyphenyl) ethyl]-4,6-quinazolinediamine [EVP-4593; Sigma–Aldrich #SML0579] at multiple timepoints (6 h, 12 h, 24 h) and compared to Time 0 (T0). EVOS M5000 Imaging System (Thermo Fisher Scientific #AMF5000) was used for cell line monitoring.