The cytotoxic activity of compounds 1 –7 was tested against A-549 human lung carcinoma cells, MDA-MB-231 human breast adenocarcinoma cells, HT-29 human colorectal carcinoma cells, and PSN1 human pancreatic adenocarcinoma cells. The four cell lines were provided by the American Type Culture Collection (ATCC): A549 from ATCC CCL-185, MDA-MB-231 from ATCC HTB-26, HT-29 from ATCC HTB-38 and PSN-1 from ATCC CRM-CRL-3211. The concentration giving half maximum inhibitory concentration (IC50) was calculated according to the procedure described in the literature [32 (link)]. Cell survival was estimated using the National Cancer Institute (NCI) algorithm [33 (link)]. Dose-response parameters were determined at three different concentrations of each one of the compounds.
Psn 1
Manufactured by American Type Culture Collection
Sourced in United States
The PSN-1 is a laboratory instrument designed for the preparation and handling of cell cultures. It provides a controlled environment for the cultivation and growth of cells, tissues, and microorganisms. The core function of the PSN-1 is to maintain the necessary temperature, humidity, and atmospheric conditions required for the optimal development and maintenance of cell cultures.
Automatically generated - may contain errors
Lab products found in correlation
Panc 1, by American Type Culture Collection (21 mentions)
Bxpc 3, by American Type Culture Collection (16 mentions)
Mia paca 2, by American Type Culture Collection (14 mentions)
Aspc 1, by American Type Culture Collection (13 mentions)
Capan 1, by American Type Culture Collection (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (7 mentions)
Sw1990, by American Type Culture Collection (6 mentions)
Hs766t, by American Type Culture Collection (6 mentions)
Fetal calf serum (fcs), by Euroclone (6 mentions)
Cfpac 1, by American Type Culture Collection (5 mentions)
Hct116, by American Type Culture Collection (5 mentions)
Mda mb 231, by American Type Culture Collection (4 mentions)
Hpaf 2, by American Type Culture Collection (4 mentions)
Hct15, by American Type Culture Collection (4 mentions)
Mia paca 2, by Thermo Fisher Scientific (3 mentions)
Panc 1, by Thermo Fisher Scientific (3 mentions)
Psn 1, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640, by Merck Group (3 mentions)
Mcf 7, by American Type Culture Collection (3 mentions)
Rpmi 1640 medium, by Euroclone (3 mentions)
42 protocols using psn 1
1
Cytotoxicity Assay of Compounds 1-7
2
Characterization of Pancreatic Cancer Cell Lines
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HPAF-II, AsPC-1, BxPC-3, CAPAN-1, CAPAN-2, CFPAC-1, Hs766T, MIAPaCa-2, Mpanc96, PANC-1, PSN-1, and SW1990 cells were obtained from ATCC (Manassas, VA, USA). Thirteen cell lines (KMP2, KMP3, KMP4, KMP5, KMP8, KP1L, KP1NL, KP2, KP3, KP3L, KP4, PH61N, and QGP-1) and normal human fibroblasts (WI-38, TIG-1, and IMR-90 cells) were obtained from JCRB cell bank (Tokyo, Japan). HPC-Y0, HPC-Y3, and HPC-Y25 were kindly given to us by Dr. Otsuji E (Kyoto Prefectural University of Medicine). PK-59 cells were obtained from RIKEN Bioresource Research Center (Ibaraki, Japan). All cell lines were cultured in RPMI-1640 medium or DMEM (Wako, Osaka, Japan) containing 10% fetal bovine serum (FBS) at 37°C with 5% CO2.
3
Cytotoxic Activity Screening of Crude Extracts
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After incubation, the broths were lyophilized and extracted with a mixture of methanol, acetone and water (1:1:0.2 v/v). The dried crude extracts were screened for cytotoxic activity (Fig. 1G in blue) against four tumour cell lines: A549 (ATCC CCL‐185) (lung carcinoma, NSCLC); HT‐29 (ATCC HTB‐38) (colon adenocarcinoma); MDA‐MB‐231 (ATCC HTB‐26) (breast adenocarcinoma); and PSN‐1 (ATCC CRL‐3211) (pancreas adenocarcinoma). All the cell lines derived from human cancer and were obtained from the American Type Culture Collection (ATCC). Cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (except PSN‐1 that was maintained in Roswell Park Memorial Institute 1640 Medium (RPMI)), supplemented with 10% fetal bovine serum (FBS), 2 mM l ‐glutamine, 100 U ml−1 penicillin and 100 U ml−1 streptomycin at 37°C, 5% CO2 and 98% humidity. For the experiments, cells were harvested from subconfluent cultures using trypsinization and resuspended in fresh medium before counting and plating. Crude extracts are resuspended with DMSO and DMEM and dispensed to culture plates containing the cells (final concentration of DMSO 1% (v/v)) to perform the cytotoxic assay. After 48 h of treatment, cytotoxic activity was measured based on the inhibition of cellular proliferation values in comparison with the control cells (Table S1 ).
4
Culturing Pancreatic Cancer Cell Lines
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Human PDAC cell lines PANC-1 (ATCC: CRL-1469), SW1990 (ATCC: CRL2172), HPAF-II (ATCC: CRL-1997), KP4 (Riken), and PSN1 (ATCC: CRM-CRL-3211) were cultured in Dulbecco’s modified eagle’s medium (DMEM) (Wisent, St-Bruno, QC) supplemented with 10% fetal bovine serum (FBS) (Wisent). Mouse cell lines 4T1 (ATCC CRL-2539) and MC38 (provided by Dr Pollak) were cultured as above. For in vitro assays, FK866 and metformin hydrochloride were purchased from Sigma-Aldrich. FK866 was suspended in DMSO and metformin in DMEM supplemented with 10% FBS. HPNE hTERT cells (ATCC) were cultured according to provider instructions.
5
Culturing Diverse Human Cell Lines
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The human cell lines AsPC-1 (CRL-1682), BxPC-3 (CRL-1687), MIA-PaCa-2 (CRM-CRL-1420), PANC-1 (CRL-1469), PSN-1 (CRL-3211) and HEK293T (CRL-1573) cells were purchased from ATCC. H6c7 cells (ECA001-FP) were purchased from Kerafast. MIA-PaCa-2, PANC-1 and HEK293T cells were maintained in DMEM with 4.5 g/l D-Glucose and GlutaMAX (Gibco), supplemented with 10% fetal calf serum (FCS, Sigma-Aldrich) and 1% Penicillin- streptomycin (P/S, Invitrogen). AsPC-1, BxPC-3, and PSN-1 were cultured in RPMI1640 (Thermo Fisher), supplemented with 10% FCS and 1% P/S. H6c7 cells were cultured in Keratinocyte serum-free medium (Thermo Fisher), supplemented with recombinant EGF and bovine pituitary extract according to the manufacturer’s instructions (Thermo Fisher), as well as 100 μg/ml Primocin (Invivogen). Cell lines were regularly checked for mycoplasma-infections by Mycoplasma PCR-detection test (Thermo-Fisher).
6
Pancreatic Cancer Cell Line Maintenance
7
Pancreatic Cancer Cell Line Cultures
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SW-1990, PSN-1, BxPC3 and PANC-1 cell lines were purchased from ATCC. MIA PaCa-2 and KP4 were purchased from Sigma-Aldrich and Accegen, respectively. MIA PaCa-2, PANC-1, SW-1990 were cultured in DMEM (Sigma-Aldrich) whereas PSN-1, BxPC3 and KP4 were cultured in RPMI1640 (Sigma-Aldrich). Both the Medias were supplemented with 10% Fetal Bovine Serum (Gibco) and 5% Penicillin-Streptomycin (Sigma-Aldrich). Cell lines were maintained at 37 °C with 5% CO2 in a cell culture incubator. Gemcitabine Hydrochloride was procured from Sigma Aldrich (catalogue number: G6423). PAK4 inhibitors (KPT-9274, catalogue number: S8444 and PF-3758309, catalogue number: S7094) were procured from Selleckchem.
8
Culturing Cancer Cell Lines
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The following cell lines were cultured in RPMI media supplemented with 10% fetal bovine serum (FBS), 2 mM l -glutamine (CORNING 25-005-CI), and 1X penicillin/streptomycin (CORNING 30-002-CI): 721.221 (Fred Hutchinson Cancer Research Center International Histocompatibility Working Group IHW00001), K562 (American Type Culture Collection (ATCC CCL-243)), Jurkat E6-1 (ATCC TIB-152), BxPC-3 (ATCC CRL-1687), CORL23 (Sigma-Aldrich 92031919), NCI-H441 (ATCC HTB-174), SW620 (ATCC CCL-227), YAPC (German Collection of Microorganisms and Cell Cultures (DSMZ) ACC 382), KP-2 (Japanese Collection of Research Bioresources Cell Bank (JCRB) JCRB0181), QGP-1 (JCRB JCRB0183), PSN-1 (ATCC CRL-3211), NCI-H2030 (ATCC CRL-5914), NCI-H358 (ATCC CRL-5807), HuCCT1 (CellBank Australia JCRB0425), and RERF-LC-Ad1 (JCRB JCRB1020). The following cell lines were cultured in DMEM media supplemented with 10% FBS, 2 mM l -glutamine, and 1X penicillin/streptomycin: SK-CO-1 (ATCC HTB-39) and HuP-T3 (DSMZ ACC 259). The following cell lines were cultured in EMEM media supplemented with 10% FBS, l -glutamine, and penicillin/streptomycin: CAL-62 (DSMZ ACC 448) and PANC1 (Laboratory of Dr Gregory Beatty at the University of Pennsylvania).
9
Cancer Cell Culture Protocols
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p53 WT cancer cells including A375, IM-9, HepG2, SKCO-1, HeLa, MCF-7, U-87-MG, LNCaP-FGC, NCI-H711, HCT116, U2OS and p53 MT cancer cells including MDA-MB-468, MOLT4, SW837, NCI-H23, P3HR1, PSN1, SKLMS1, SKLU1, SK-UT-1 and SNU-16 lines were procured from ATCC (VA, USA). HEK293T cells were procured from Cell BioLabs Inc. (San Diego, CA) and HEK293FT cells were procured by Thermo Fisher Scientific (MA, USA). Briefly, the cells were cultured in monolayer in respective growth mediums (Dulbecco's modified Eagle's medium (DMEM) or RPMI 1640) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS) and antibiotics (1% penicillin/streptomycin) and incubated under normoxic conditions at 37°C in a humidified atmosphere of 95% air and 5% CO2.
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p53 WT cancer cells including A375, IM-9, HepG2, SKCO-1, HeLa, MCF-7, U-87-MG, LNCaP-FGC, NCI-H711, HCT116, U2OS and p53 MT cancer cells including MDA-MB-468, MOLT4, SW837, NCI-H23, P3HR1, PSN1, SKLMS1, SKLU1, SK-UT-1 and SNU-16 lines were procured from ATCC (VA, USA). HEK293T cells were procured from Cell BioLabs Inc. (San Diego, CA) and HEK293FT cells were procured by Thermo Fisher Scientific (MA, USA). Briefly, the cells were cultured in monolayer in respective growth mediums (Dulbecco's modified Eagle's medium (DMEM) or RPMI 1640) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS) and antibiotics (1% penicillin/streptomycin) and incubated under normoxic conditions at 37°C in a humidified atmosphere of 95% air and 5% CO2.
10
Pancreatic Cancer Cell Line Culturing
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The MIT Animal Care and Use Committees reviewed and approved all animal studies and procedures. NOD/SCID/IL2Rγ-null mice (Jackson Laboratory) were used throughout the study. Mice were housed on a 12-h light/12-h dark cycle at ~21 °C and 40–60% humidity. The human pancreatic adenocarcinoma (PDAC) cell lines AsPC1, BxPC3, PANC1, and MIAPaCa2 and HEK 293FT cells were purchased from American Type Cell Culture (ATCC), where they were tested and authenticated; PSN1 and CFPAC1 were gifts from the Koch Institute cell line repository, and they were originally tested and authenticated from ATCC. The human CAF cell line hT1 and hM1 were published previously59 (link). All cells were cultured in media supplemented with 10% fetal bovine serum (FBS, Invitrogen) at 37 °C in a 5% CO2 incubator. AsPC1 was cultured in RPMI medium 1640 (ThermoFisher); 293FT, BxPC3, PANC1, MIAPaCa2 and PSN1 were cultured in Dulbecco’s modified Eagle’s medium (DMEM, ThermoFisher); CFPAC1 was cultured in Iscove’s modified Dulbecco’s Medium (ThermoFisher).
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