Human OC cell lines, HeyA8 (received from Dr. Ernst Lengyel, RRID:CVCL_8878), OVCAR5 (obtained from the Developmental Therapeutics Program at the National Cancer Institute, RRID:CVCL_1628), were cultured in RPMI 1640 medium (Cell gro #10–040 CM), supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Sigma-Aldrich # 14009C), 1% L-glutamine (Mediatech Inc), and 1% penicillin/streptomycin (Mediatech Inc); PEO1 (RRID:CVCL_2686) and PEO4 (RRID:CVCL_2690) cells both obtained from Sigma-Aldrich were cultured in the above medium and supplemented with 2 mM sodium pyruvate (Mediatech Inc). 293T WT, 293T Dicer k.o. cells [Clone #2–20; (28 (link))], HeLa control (RRID:CVCL_0030) and HeLa Ago2 k.o. were a kind gift from Dr. Sarah Gallois-Montbrun (29 (link)), 293T Ago2 k.o. cells were provided by Dr. Klaas Mulder (30 (link)), and OC cell lines A2780 (RRID:CVCL_4862) and A2780R (27 ) were all grown in DMEM medium (Gibco #12430054) supplemented with 10% FBS, 1% L-glutamine and 1% penicillin/streptomycin. All cell lines were authenticated by STR profiling at IDEXX BioAnalytics. Cells were used within the first 3–8 passages following thawing, periodically tested for mycoplasma using PlasmoTest (Invitrogen) and were last tested in April 2021.
Plasmotest
Manufactured by Thermo Fisher Scientific
PlasmoTest is a rapid diagnostic test designed for the detection of Plasmodium falciparum, the most prevalent and deadly form of malaria. The core function of PlasmoTest is to provide a qualitative, rapid and visual detection of P. falciparum antigens in whole blood samples.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Corning (3 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Corning (2 mentions)
Rpmi 1640 medium, by Corning (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Sodium pyruvate, by Corning (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Dmem medium, by Corning (1 mentions)
Mccoy s 5a medium, by American Type Culture Collection (1 mentions)
Serum plus 2 medium supplement, by Merck Group (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Hek293t, by American Type Culture Collection (1 mentions)
Hek293, by American Type Culture Collection (1 mentions)
Flp in t rex 293 cell line, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
4 protocols using plasmotest
1
Culturing Human OC Cell Lines
2
Culturing and Maintaining Cell Lines
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293T cells were cultured in DMEM medium (Corning #10–013-CM) with 10% Serum Plus II Medium Supplement (Sigma-Aldrich #14009C). Generation of MCF7 CD95 k.o. clone was described in [16 (link)]. MCF7 CD95 k.o. cells were cultured in RPMI 1640 medium (Corning #10–040 CM) supplemented with 10% Serum Plus II Medium Supplement (Sigma-Aldrich #14009C) and 1% penicillin/streptomycin (Mediatech Inc). The following HCT116 cells were purchased from the Korean Collection for Type Cultures (KCTC): HCT116 wild-type (KCTC, cat #HC19023), HCT116 Drosha k.o. clone #40 (KCTC, cat #HC19020), HCT116 Dicer k.o. clone #43 (KCTC, cat #HC19023). The HCT116 Ago2 k.o. cells [48 (link)] were a kind gift from Joshua T. Mendell (UT Southwestern). The HCT116 Ago1 k.o. and HCT116 Ago 1/2/3 k.o. cells [26 (link)] were provided by David Corey (UT Southwestern). All HCT116 cells and knock-out clones were cultured in McCoy’s 5A medium (ATCC #30–2007) with 10% Serum Plus II Medium Supplement (Sigma-Aldrich #14009C). Cells were cultured at 37 °C, 5% CO2 were tested for mycoplasma using PlasmoTest (Invitrogen). Before use the Serum Plus II Medium Supplement was heat inactivated by incubation at 56 °C for 30 min.
3
Cell Culture Conditions for Various Cancer Cell Lines
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HeyA8 (RRID:CVCL_8878 ) and HeyA8 CD95 knock-out cells, HCT116 (ATCC #CCL-247; RRID:CVCL_0291 ) and HCT116 Drosha knock-out and Dicer knock-out cells, MCF-7 cells (ATCC #HTB-22; RRID:CVCL_0031 ), and 293T (ATCC #CRL-3216; RRID:CVCL_0063 ) cells were cultured as described previously (Putzbach et al., 2017 (link)). The MCF-7 CD95 knock-out and deletion cells were cultured in RPMI 1640 medium (Cellgro #10–040 CM), 10% heat-inactivated FBS (Sigma-Aldrich), 1% L-glutamine (Mediatech Inc), and 1% penicillin/streptomycin (Mediatech Inc). H460 (ATCC #HTB-177; RRID:CVCL_0459 ) cells were cultured in RPMI1640 medium (Cellgro Cat#10–040) supplemented with 10% FBS (Sigma Cat#14009C) and 1% L-glutamine (Corning Cat#25–005). 3LL cells (ATCC #CRL-1642; RRID:CVCL_4358 ) were cultured in DMEM medium (Gibco Cat#12430054) supplemented with 10% FBS and 1% L-glutamine. Mouse hepatocellular carcinoma cells M565 cells were described previously (Ceppi et al., 2014 (link)) and cultured in DMEM/F12 (Gibco Cat#11330) supplemented with 10% FBS, 1% L-glutamine and ITS (Corning #25–800-CR). All cell lines were authenticated using STR profiling and tested monthly for mycoplasm using PlasmoTest (Invitrogen).
4
Cultivation of HEK293 and Engineered Derivatives
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HEK293 and HEK293T from the American Type Culture Collection (ATCC) were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Life Technologies) with 10% FCS (Life Technologies) and antibiotics (Life Technologies). The EGFP T-REx 293 cell line used for the mammalian screening was obtained by modifying the Flp-In™ T-REx™ 293 Cell Line (Invitrogen) for the expression of inducible EGFP according to the manufacturer’s protocol. The cell lines were verified for the absence of mycoplasma contamination (PlasmoTest, Invitrogen).
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