Human pancreatic cancer cell lines, AsPC-1, Hs766T, MIA PaCa-2, PANC-1, and Capan-1, were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The human immortalized pancreas epithelial cell line, hTERT-HPNE, was also purchased from the ATCC. Luciferase expressing Hs766T cells (Hs766T (+Luc)) were generated as previously described. AsPC-1 pancreatic cancer cells were cultured in RPMI-1640 Medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS) (MilliporeSigma, St. Louis, MO, USA). Hs766T, Hs766T (+Luc), MIA PaCa-2, and PANC-1 pancreatic cancer cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Thermo Fisher Scientific, Waltham, MA) supplemented with 10% FBS. Capan-1 cells were cultured in Iscove’s Modified Dulbecco’s medium (IMDM) (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS. hTERT-HPNE cells were cultured in 75% DMEM without glucose (Thermo Fisher Scientific, Waltham, MA, USA) and 25% Medium M3 Base (Incell Corporation, San Antonio, TX, USA) supplemented with 5% FBS, human recombinant EGF (10 ng/mL) (Thermo Fisher Scientific, Waltham, MA, USA), D-glucose (5.5 mM) (MilliporeSigma, St. Louis, MO, USA), and puromycin (750 ng/mL) (MilliporeSigma, St. Louis, MO, USA).
Medium m3 base
Manufactured by INCELL
Sourced in United States
The Medium M3 Base is a laboratory equipment designed to provide a stable platform for various experimental setups. It features a sturdy construction and a flat surface to accommodate various apparatus and instruments. The core function of the Medium M3 Base is to offer a reliable and secure foundation for experimental work.
Automatically generated - may contain errors
Lab products found in correlation
Htert hpne, by American Type Culture Collection (11 mentions)
Panc 1, by American Type Culture Collection (10 mentions)
Mia paca 2, by American Type Culture Collection (7 mentions)
Aspc 1, by American Type Culture Collection (5 mentions)
Sw620, by American Type Culture Collection (5 mentions)
Htert hpne, by Thermo Fisher Scientific (4 mentions)
Htert hpne, by Merck Group (4 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions)
D glucose, by Merck Group (4 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (4 mentions)
Rpmi 1640, by Merck Group (4 mentions)
Hct116, by American Type Culture Collection (4 mentions)
Recombinant human egf, by Thermo Fisher Scientific (3 mentions)
Dmem without glucose, by Merck Group (3 mentions)
Bxpc 3, by American Type Culture Collection (3 mentions)
Sw1990, by American Type Culture Collection (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Capan 2, by American Type Culture Collection (3 mentions)
Ncm460, by INCELL (3 mentions)
Sw480, by American Type Culture Collection (3 mentions)
33 protocols using medium m3 base
1
Culturing Human Pancreatic Cell Lines
2
Pancreatic Cancer Cell Line Cultivation
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Pancreatic cell lines (PANC-1, BxPC3, CAPAN-2, MiaPaca2, hTERT-HPNE, PL-45 and Su.86.86) were obtained from the American Tissue Type Collection and cultured as previously described (18 (link)). Specifically, PANC-1, MiaPaca2,CAPAN-2 and PL-45 were cultured in DMEM high glucose (MilliporeSigma) with 10% Fetal Bovine Serum (FBS; Euroclone); Su.86.86 and BxPC3 cells were cultured in RPMI-1640 (MilliporeSigma)/10% FBS; hTERT-HPNE were cultured in 75% DMEM (MilliporeSigma) (with 2 mM L-glu and 1.5 g/l sodium bicarbonate, both from MilloporeSigma)/25% Medium M3 Base (Incell Corp.) with 5% FBS, 10 ng/ml human recombinant EGF (Thermo Fisher Scientific, Inc.), 5.5 mM D-glucose (1 g/l; MilliporeSigma) and 750 ng/ml puromycin (Thermo Fisher Scientific, Inc.). The transfections of plasmids were performed with FuGENE HD (Promega Corporation) according to the manufacturer's protocol. Short hairpin (sh)RNA for COUP-TFII (shNR2F2) and negative control shRNA (shNEG) are described in (18 (link)); shNR2F2 covers the same target sequence of a small interfering (si)RNA for COUP-TFII (Hs_NR2F2_6, cat. no. s103649065; Qiagen GmbH). COUP-TFII_V2-GFP (cat. no. RG226609) and COUP-TFII_V2 (cat. no. 4453629) plasmids were from OriGene Technologies, Inc.
3
Characterization of Pancreatic Cancer Cell Lines
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The human PDAC cell lines PANC-1, MiaPaCa-2, AsPC-1, BxPC-3, SW1990 and immortalized human pancreas duct epithelial cell line hTERT-HPNE cells were purchased from the American Type Culture Collection (ATCC). FG were described previously [28 (link)]. All of these cancer cell lines were maintained in plastic flasks as adherent monolayers in Eagle’s minimal essential medium supplemented with 10% fetal bovine serum, sodium pyruvate, nonessential amino acids, L-glutamine, and a vitamin solution (Flow Laboratories). hTERT-HPNE cells were cultured in a mixture of Dulbecco’s Modified Eagle’s Medium without glucose (Sigmaich-Aldrich, Cat. No. D-5030) and Medium M3 Base (Incell Corp, Cat. No. M300F-500) (3:1 ratio) with 2 mML-glutamine adjusted to 1.5 g/L sodium bicarbonate and supplemented with 5% FBS, 10 ng/ml human recombinant EGF, 5.5 mM D-glucose (1 g/L) and 750 ng/mL puromycin. The cell lines were obtained directly from ATCC that performs cell line characterizations or authentication by the short tandem repeat profiling and passaged in our laboratory for fewer than 6 months after receipt.
The following drugs were used with an indicated concentration in the experiments: MST1/2 activator okadaic acid (OA) and de-methylating agent 5-aza-2′-deoxycytidine (5-aza) were purchased from Sigma [3 (link), 29 (link)].
The following drugs were used with an indicated concentration in the experiments: MST1/2 activator okadaic acid (OA) and de-methylating agent 5-aza-2′-deoxycytidine (5-aza) were purchased from Sigma [3 (link), 29 (link)].
4
HPNE and Pancreatic Cancer Cell Lines
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The hTERT-immortalized Human Pancreatic Nestin-Expressing cells (HPNE) were purchased from the American Type Culture Collection (ATCC, Molsheim, France). HPNE were grown in 75% DMEM without glucose (Sigma, Cat no. D-503, St. Louis, MO, USA), 25% Medium M3 Base (Incell Corp., San Antonio, TX, USA, Cat no. M300F-500), which were completed with 10% fetal bovine serum (Cat no. P30-3031, PAN Biotech, Aidenbach, Germany), 5.5 mM glucose and 750 ng/mL puromycin. Colo357, BxPC-3, AsPC-1, MIA PaCa-2, and PANC-1 cells were kindly provided by Prof. Anna Trauzold (Institute of Experimental Cancer Research, Kiel University, Kiel, Germany). These cell lines were grown in RPMI-1640 (Sigma-Aldrich, Cat no. R8758), supplemented with 10% fetal bovine serum (Cat no. P30-3031, PAN Biotech), 1 mM sodium pyruvate (Gibco, Waltham, MA, USA), and 1% Glutamax (Gibco). Cells were grown at 37 °C, 95% humidity, 5% CO2, and passaged with trypsin-EDTA 0.25% (Sigma, Saint-Quentin-Fallavier, France) when cells reached a confluency of 70–80%. Cell cultures were not used for more than 20 passages.
5
Pancreatic Cancer Cell Lines Mitophagy
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The human pancreatic cancer cell lines (PANC-1, BxPC-3 and HPAC) and normal pancreatic ductal epithelial cell line (hTERT-HPNE) were purchased from the American Type Culture Collection (ATCC). The PANC-1, BxPC-3 cells and HPAC cells were all cultured in RPMI-1640 medium (Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (FBS; HyClone; GE Healthcare Life Sciences), 1% L-glutamine and 0.5% gentamycin (Sigma-Aldrich; Merck KGaA) at 37°C in an incubator with 5% CO2. The hTERT-HPNE cells were cultured in medium containing three volumes of glucose-free DMEM, one volume of Medium M3 base (InCell), 5% FBS, 5.5 mM glucose, 10 ng/ml human recombinant EGF and 50 µg/ml gentamicin (27 (link)). To activate mitochondrial mitophagy, cells were treated with 5 µM FCCP (Selleck Chemicals) for 2 h at 37°C prior to treatment.
6
Pancreatic Cancer Cell Culture Protocol
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The KP3 human pancreatic adenocarcinoma cell line expressing luciferase reporter gene was obtained from the Japanese Collection of Research Bioresources Cell Bank (Tokyo, Japan). Panc-1 and BxPC3 cell lines and the normal pancreatic ductal epithelial cell line hTERT-HPNE were obtained from the American Type Culture Collection (Manassas, VA). KP3, Panc-1, and BxPc3 cells were cultured in Dulbecco’s modified Eagle medium (DMEM, 4.5 g/L glucose; Thermo Fisher Scientific, Waltham, MA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. hTERT-HPNE cells were cultured with a mixture of 75% DMEM (no glucose; Thermo Fisher Scientific), and 25% Medium M3 Base (InCell, Frisco, TX) supplemented with 5% FBS, 10 ng/mL human recombinant epidermal growth factor, 1g/L glucose, and 750 ng/mL puromycin. All cells were maintained under humidified conditions (5% CO2, 37°C).
7
Chronic Cadmium Exposure in Epithelial Cells
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We used two human non-cancer epithelial cell lines: one from breast (MCF10A), and one from pancreas (hTERT-HPNE). MCF10A were purchased from ATCC® (CRL-10317™) and were exposed with 2.5 µM Cd 2+ for 40 weeks according to previously published data (Benbrahim-Tallaa et al. 2009) (link). hTERT-HPNE were purchased by ATCC® (CRL-4023™) and exposed with 1 µM Cd 2+ for 30 weeks (Qu et al. 2012 (link)). MCF10A were cultured in Dulbecco's Modified Eagle's Medium/Nutrient Mixture F-12 (DMEM/F-12) supplemented with 10% Fetal Calf Serum (FCS), hydroxycortisone (50 ng/mL), insulin (0.01 mg/mL), human recombinant EGF (20 ng/mL), and cholera toxin (100 ng/mL). hTERT-HPNE were cultured in base medium: 75% DMEM without glucose (Sigma) and 25% Medium M3 Base (Incell Corp) with 5% FCS. Complete growth medium was supplemented with the following components: human recombinant EGF (10 ng/mL), D-glucose (1 g/L), puromycin (750 ng/mL), l-glutamine (0.3 g/L), and sodium bicarbonate (1.5 g/L). Cells were trypsinized once a week using trypsin-EDTA (Sigma-Aldrich, Inc.) and incubated at 37 °C in a humidified atmosphere containing 5% CO 2 .
8
Synthesis and Characterization of PEG-PGlu Copolymers
9
Cultivation of Pancreatic Cell Lines
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Human pancreatic adenocarcinoma cell lines (PANC-1. MIA PaCa-2 and AsPC-1) and were acquired from the Cell Resource Centre of Peking Union Medical College (Beijing, China). Pancreatic duct normal cell lines (hTERT-HPNE) were purchased from American Type Culture Collection (ATCC). PANC-1 and MIA Paca-2 were cultivated in Dulbecco's modified Eagle's medium (DMEM, Cell Resource Centre of Peking Union Medical College, Beijing, China), while AsPC-1 was in RPMI-1640 (Cell Resource Centre of Peking Union Medical College, Beijing, China), and both cell lines were supplemented with 10% fetal bovine serum (FBS; product number: 35-010-CV; Corning, NY, USA) and penicillin and streptomycin (100 U/ml; catalogue number: 10378-016; Thermo Fisher Scientific, San Jose, USA) in 75-cm2 flasks in a 37°C incubator with 5% CO2. hTERT-HPNE was cultivated in 75% DMEM without glucose (Sigma Cat#. D-5030 with additional 2 mM L-glutamine and 1.5 g/L sodium bicarbonate) and 25% Medium M3 Base (Incell Corp. Cat# M300F- 500), with a supplementary of 5% FBS, 15 ng/ml human recombinant EGF, 5.5mM D-glucose (1g/L) and 2μg/ml puromycin. The cells were passaged into 3 flasks when adherent cells accounted for 75% of the total area. Recombinant p53 protein was purchased from Origene (Catalog: TP710022) and Cell culture experiments were used to verify the predictions.
10
Culturing Pancreatic Cancer and Nestin-Expressing Cell Lines
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Pancreatic cancer cell-line PANC-1 was procured from ATCC (CRL-1469; Manassas, VA, USA) and cultured in Dulbecco’s Modified Eagle’s Medium (30–2002, ATCC) supplemented with 10% FBS (Sigma, St.Louis, MI, USA). MIA PaCa-2 was obtained from ATCC (CRM-CRL-1420) and cultured in Dulbecco’s Modified Eagle’s Medium (30–2002, ATCC) supplemented with 10% FBS. AsPC-1 was obtained from ATCC (CRL-1682), and cultured in an RPMI-1640 medium (30–2001, ATCC) supplemented with 10% FBS (Sigma). Capan-1 was obtained from ATCC (HTB-79), and cultured in an Iscove’s Modified Dulbecco’s Medium (30–2005, ATCC) supplemented with 20% FBS (Sigma).
A human pancreatic nestin-expressing cell line,27 (link) hTERT-HPNE, was obtained from ATCC and cultured in amixture of 75% DMEM without glucose (Cat#. D-5030; Sigma, with additional 2 mM L-glutamine and 1.5 g/L sodium bicarbonate) and 25% Medium M3 Base (Cat# M300F-500; Incell Corp., San Antonio, TX, USA) supplemented with 10% FBS (Sigma). All the cells were cultured at 37°C in 5% CO2.
A human pancreatic nestin-expressing cell line,27 (link) hTERT-HPNE, was obtained from ATCC and cultured in amixture of 75% DMEM without glucose (Cat#. D-5030; Sigma, with additional 2 mM L-glutamine and 1.5 g/L sodium bicarbonate) and 25% Medium M3 Base (Cat# M300F-500; Incell Corp., San Antonio, TX, USA) supplemented with 10% FBS (Sigma). All the cells were cultured at 37°C in 5% CO2.
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