The human pancreatic cancer cell lines MIA PaCa-2, AsPC-1, PANC-1 and BxPC-3 were purchased from American Type Culture Collection (CRL-1420TM, CRL-1682, CRL-1469 and CRL-1687, respectively, ATCC; Rockville, MD, USA) and banked at Centre Paul Strauss. Cells were cultured at 37 °C in a humidified atmosphere containing 5% CO2 and 95% air. PANC-1 was cultured in Dulbecco’s modified Eagle’s medium (DMEM; PAN Biotech GmbH, Aidenbach, Germany) supplemented with 10% fetal bovine serum (FBS, PAN Biotech GmbH) and 1% of a solution of penicillin (10000 IU/mL) and streptomycin (10 mg/mL) (PAN Biotech GmbH). MIA PaCa-2 was cultured in the same conditions with 1% of 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, 1 mM, PAN Biotech GmbH), 1% of sodium pyruvate (10 mM, PAN Biotech GmbH) and 1% of non-essential amino acids (NEAA, PAN Biotech GmbH). AsPC-1 and BxPC-3 were cultured in Roswell Park Memorial Institute medium (RPMI; PAN Biotech GmbH) supplemented with 10% FBS, and 1% of a solution of penicillin (10000 IU/mL) and streptomycin (10 mg/mL). Subconfluent cell monolayers were trypsinized once a week using 0.5% trypsin containing 2% EDTA (PAN Biotech GmbH) and plated at passage ratios between 0.25:10 to 1:10, according to the cell line or used directly for study after enumeration determined with a Countess® Cell Counter (Countess, Invitrogen, Carlsbad, CA, USA).
Crl 1687
Manufactured by American Type Culture Collection
Sourced in United States
The CRL-1687 is a cell line derived from the lung tissue of a human individual. It is maintained in culture and can be used for various research applications, such as studying cellular processes or testing the effects of different treatments or conditions on cell behavior.
Automatically generated - may contain errors
Lab products found in correlation
Panc 1, by American Type Culture Collection (4 mentions)
Crl 1469, by American Type Culture Collection (3 mentions)
Bxpc 3, by American Type Culture Collection (3 mentions)
Aspc 1, by American Type Culture Collection (2 mentions)
Sodium pyruvate, by PAN Biotech (1 mentions)
Countess cell counter, by Thermo Fisher Scientific (1 mentions)
Crl 1682, by American Type Culture Collection (1 mentions)
Panc 1, by PAN Biotech (1 mentions)
Mia paca 2, by PAN Biotech (1 mentions)
4 2 hydroxyethyl 1 piperazineethanesulfonic acid hepes, by PAN Biotech (1 mentions)
Bxpc 3, by PAN Biotech (1 mentions)
Non essential amino acids (neaa), by PAN Biotech (1 mentions)
Mia paca 2, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by PAN Biotech (1 mentions)
Penicillin, by PAN Biotech (1 mentions)
Streptomycin, by PAN Biotech (1 mentions)
Ccl 247, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Ultra low attachment plates, by Thermo Fisher Scientific (1 mentions)
Htb 14, by American Type Culture Collection (1 mentions)
9 protocols using crl 1687
1
Culturing Pancreatic Cancer Cell Lines
2
Maintenance and Spheroid Formation of Cancer Cell Lines
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HCT116 (human colon carcinoma, (ATCC® CCL‐247™)), BxPC3 (human pancreas adenocarcinoma (ATCC® CRL‐1687™), and U87 (human glioblastoma (ATCC® HTB‐14™)) cell lines were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco), supplemented with 10% fetal bovine serum (FBS; Millipore) and penicillin/streptomycin (Life Technologies), and cultured at 37°C, 5% CO2, and 95% humidity. For spheroid formation, 10,000 cells were plated in ultra‐low attachment plates (Fisher Scientific), centrifuged at 850 g for 10 min, and incubated for 3 days until treatment, with medium change every 3 days. The absence of contamination with mycoplasma was ensured by regular testing using the PCR Mycoplasma Test Kit I/C (PromoKine PK‐CA91‐1048). Cell authentication is ensured by the supplier (ATCC).
3
Cadmium Chloride Cytotoxicity in Pancreatic Cells
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The cell lines are obtained from the American Type Culture Collection (ATCC, Manassas, VA). Control pancreas cells [hTERT-HPNE (“human pancreatic Nestin-expressing” cells or HPNE; ATCC® CRL-4023™, control pancreatic cells)] and tumour [AsPC-1 (ATCC® CRL-1682™, pancreatic tumour cells), Panc-1 (ATCC #CRL-1469™, Pancreas ductal epithelioid carcinoma), Panc-10.05 (ATCC #CRL-2547™, Pancreatic epithelial adenocarcinoma), and BxPC-3 (ATCC #CRL-1687™, Pancreatic adenocarcinoma)] were grown and maintained as described in the ATCC-suggested protocols. Unless otherwise specified, cells were grown in their defined optimum growth media. For a parallel set of LC50 assays, cells were grown in a minimal media of MEM plus 1% foetal bovine serum (FBS). The cells were split in 6-well plates at 3–5 day intervals depending on confluence and cells were grown to 80% confluency in preparation for 14 days treatment with cadmium chloride (CdCl2; 50 μM), based on our previously published data (Djordjevic et al 2019 (link)).
4
Cytotoxicity Evaluation of Irinotecan Nanoparticles
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The CT-26 murine colon carcinoma (CT-26; ATCC, CRL-2638) and BxPC3 human pancreatic carcinoma (BxPC3, ATCC® CRL1687™) were cultured in RPMI-1640 media supplemented with 10% FBS, 50 U mL -1 penicillin, 50 µg mL -1 streptomycin, 1% L-glutamine, at 37 °C in 5% CO 2 . Cells were routinely grown in 75 cm 2 canted-neck tissue culture flasks and passaged twice a week using trypsin/EDTA at 80% confluence.
CT-26 or BxPC3 cells were seeded in 96-well plates at 4000 cells per well and incubated with different concentrations of irinotecan or its NP (0.1-100 μM irinotecan concentration) in complete media for 72 h. Cells were also treated with blank NP formulations (P-NP and T-NP) at the polymer concentrations equivalent to drug NP formulations (P-NP-Ir AND T-NP-Ir). Cytotoxicity was examined by MTT assay. Briefly, at the end of the incubation period, the media was removed and replaced with 120 μL of MTT solution at a final concentration of 0.5 mg mL -1 . Cells were incubated for 3 h at 37 °C and 5% CO 2 . At the end of the incubation, formazan was dissolved in 200 μL of DMSO and the plate was read at 570 nm in an FLUO star OPTIMA plate reader (BMG Labtech) and the results were expressed as the percentage cell survival (mean ± SD) and calculated using the following equation: % cell survival = (A570 nm of treated cells/A570 nm of untreated control cells) × 100.
CT-26 or BxPC3 cells were seeded in 96-well plates at 4000 cells per well and incubated with different concentrations of irinotecan or its NP (0.1-100 μM irinotecan concentration) in complete media for 72 h. Cells were also treated with blank NP formulations (P-NP and T-NP) at the polymer concentrations equivalent to drug NP formulations (P-NP-Ir AND T-NP-Ir). Cytotoxicity was examined by MTT assay. Briefly, at the end of the incubation period, the media was removed and replaced with 120 μL of MTT solution at a final concentration of 0.5 mg mL -1 . Cells were incubated for 3 h at 37 °C and 5% CO 2 . At the end of the incubation, formazan was dissolved in 200 μL of DMSO and the plate was read at 570 nm in an FLUO star OPTIMA plate reader (BMG Labtech) and the results were expressed as the percentage cell survival (mean ± SD) and calculated using the following equation: % cell survival = (A570 nm of treated cells/A570 nm of untreated control cells) × 100.
5
Cell Viability Assay for Cancer Cells
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Human colon cancer cells (HT29), human melanoma cells (MDA-MB-435), human lymphoblast cells (ARH77) and human pancreatic cancer cells (CRL1687) were from ATCC and maintained in RPMI-1640 with L-Glutamine and Phenol Red, supplemented with 10% (v/v) heat-inactivated Fetal Calf Serum (FCS), 15mM HEPES, 1% (v/v) penicillin-streptomycin, and 1% L-glutamine. Cell viability test was done by MTT test according to manufacturer's protocol. Thiazolyl blue tetra-zolium bromide is from Sigma Aldrich (St Louis, USA).
6
Culturing Cancer and Normal Cell Lines
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Cancer cell lines: BxPC-3 (pancreas adenocarcinoma, ATCC® CRL-1687TM), and HT-29 (colorectal adenocarcinoma, ATCC® HTB-38TM) and normal cell line—WI-38 (human lung fibroblasts, ATCC® CCL-75TM) were obtained from American Type Culture Collection (ATCC, Rockville, MD, USA). BxPC-3 cells were grown in RPMI-1640 medium supplemented with 10% (v/v) FBS and 1% (v/v) of both antibiotics (streptomycin and penicillin). For the HT-29 cell line, RPMI-1640 medium supplemented with 10% (v/v) FBS and 1% (v/v) of both antibiotics and 1% MEM nonessential amino acids was used to ensure proper cell growth. WI-38 cells were grown in MEM medium supplemented with 10% (v/v) FBS, L-Glutamine, 25 mM Hepes, and 1% penicillin–streptomycin. MycoBlueTM Mycoplasma Detector kit (Vazyme Biotech, Nanjing, China) was used at least every month for the control of mycoplasma contamination in the cell cultures.
Cells were grown at 37 °C in a humidified atmosphere of 5% CO2 in the air. The culture medium was changed every 24–48 h. Subculture was performed using 0.25% trypsin/EDTA after cells reached confluence.
Cells were grown at 37 °C in a humidified atmosphere of 5% CO2 in the air. The culture medium was changed every 24–48 h. Subculture was performed using 0.25% trypsin/EDTA after cells reached confluence.
7
Cultivation and Characterization of Cell Lines
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Cancer cell lines: BxPC-3 (pancreas adenocarcinoma, ATCC® CRL-1687TM), HCT-116 (colorectal carcinoma, ATCC® CCL-247™), PC-3 (prostate cancer, ATCC® CRL-1435TM), and normal cell lines: L929 (mouse fibroblast, ATCC® CCL-1™) and WI-38 (human lung fibroblasts, ATCC® CCL-75TM) were obtained from American Type Culture Collection (ATCC, Rockville, MD, USA). Compositions of culture media used to cultivate cells are presented in Table 8 .
Cells were grown at 37 °C in a humidified atmosphere of 5% CO2 in the air. The culture medium was changed every 24–48 h. Subculture was performed using 0.25% trypsin/EDTA after cells reached confluence.
Cells were grown at 37 °C in a humidified atmosphere of 5% CO2 in the air. The culture medium was changed every 24–48 h. Subculture was performed using 0.25% trypsin/EDTA after cells reached confluence.
8
Hypoxia Effects on Pancreatic Cancer Cells
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Human pancreatic cancer cell lines (BxPC3; CRL-1687, PANC-1; CRL-1469) were purchased from the American Tissue Culture Collection (ATCC, USA). PANC-1 cells were maintained in DMEM with 10% FBS, 100 U/ml penicillin, and 100 µg/ml streptomycin. BxPC3 cells were maintained in RPMI-1640 with 10% FBS, 100 U/ml penicillin, and 100 µg/ml streptomycin. PANC-1 (3x105 cells per flask) or BxPC3 (1x106 cells per flask) were seeded in 75cm2 cell culture flasks in 10ml of medium and were incubated under normoxic conditions (20.9% O2, 5% CO2) for 6 days or 4 days, respectively. The medium was then changed, and the cells incubated in normoxia for a further 24h before being cultured for a further 24h under either normoxic or hypoxic (0.5% O2, 5% CO2) conditions.
9
Culturing Human Pancreatic Cell Lines
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Human pancreatic cancer cell lines (BxPC3; CRL-1687, PANC-1; CRL-1469) were purchased from the American Tissue Culture Collection (ATCC, USA). The immortalized epithelial cell line (HPDE) that derived from normal human pancreatic duct was provided by Dr. Ming-Sound Tsao (Ontario Cancer Institute, Canada) (Furukawa et al., 1996 (link); Makawita et al., 2011 ; Pramanik et al., 2011 (link); Radulovich et al., 2008 (link)). PANC-1 cells were maintained in DMEM with 10% FBS, 100 U/ml penicillin, and 100 mg/mL streptomycin. BxPC3 cells were maintained in RPMI-1640 with 10% FBS, 100 U/ml penicillin, and 100 mg/mL streptomycin. HPDE cells were maintained in Keratinocyte-SFM media with bovine pituitary extract (BPE) and human recombinant EGF. All cells were cultured in a 37°C incubator with 5% CO2 and a humidified atmosphere.
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