The human pancreatic cancer cell lines BxPC-3 (cat# CRL-1687), AsPC-1 (CRL-1682), CFPAC-1 (CRL-1918), and PANC-1 (CRL-1469) were obtained from the American Type Culture Collection (ATCC) (Manassas, VA, USA). Both BxPC-3 and AsPC-1 cells were cultured in RPMI 1640 medium (SH30255.01; HyClone, Logan, UT, USA) with 1.0% sodium pyruvate and 10% FBS (100–106; Gemini Bio Products, Sacramento, CA, USA). The CFPAC-1 cells were cultured in IMDM (30–20065; ATCC) with 10% FBS. Each cell line's identity was confirmed by the Genetic Resources Core Facility of Johns Hopkins University (Baltimore, MD, USA). Additionally, HEK293 cell lines that stably express the human DUOX2 gene alone or with human DUOXA2 were kindly provided by Dr. William M. Nauseef (University of Iowa, Iowa City, IA, USA) and were maintained as described [63 (link)]. To establish starvation conditions before each experiment, cells were cultured overnight in the same medium without FBS. Starvation conditions were used because DUOX2 induction by IFN-γ is stronger after serum starvation, as noted previously [11 (link), 12 (link)]. In all cases, cells were cultured in a humidified incubator at 37°C in an atmosphere of 5% CO2 in air. We previously found that the basal levels of all Nox homologues (Nox1-5 and DUOX1-2) were negligible in BxPC-3, AsPC-1, and CFPAC-1 cells, as determined by real-time RT-PCR [11 (link)].
Cfpac 1
Manufactured by American Type Culture Collection
Sourced in United States, China, Japan, Italy, United Kingdom
CFPAC-1 is a cell line derived from a primary human pancreatic adenocarcinoma. It is a widely used model for the study of pancreatic cancer.
Automatically generated - may contain errors
Lab products found in correlation
Panc 1, by American Type Culture Collection (211 mentions)
Bxpc 3, by American Type Culture Collection (190 mentions)
Aspc 1, by American Type Culture Collection (150 mentions)
Mia paca 2, by American Type Culture Collection (142 mentions)
Sw1990, by American Type Culture Collection (102 mentions)
Capan 2, by American Type Culture Collection (83 mentions)
Capan 1, by American Type Culture Collection (74 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (68 mentions)
Hpaf 2, by American Type Culture Collection (36 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (29 mentions)
Cfpac 1, by Thermo Fisher Scientific (27 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (25 mentions)
Hs766t, by American Type Culture Collection (22 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (20 mentions)
Htert hpne, by American Type Culture Collection (20 mentions)
Panc 1, by Thermo Fisher Scientific (17 mentions)
Su 86, by American Type Culture Collection (16 mentions)
Penicillin, by Thermo Fisher Scientific (16 mentions)
Streptomycin, by Thermo Fisher Scientific (14 mentions)
Bxpc 3, by Thermo Fisher Scientific (14 mentions)
296 protocols using cfpac 1
1
Culturing Human Pancreatic Cancer Cell Lines
2
Evaluating Pancreatic Cancer Cell Migration
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PC cell lines AsPC-1, PANC-1, CFPAC-1, MIA PaCa-2, BxPC-3 and normal pancreatic ductal epithelial cell line HPNE were all purchased from the ATCC (Manassas, VA, USA). All cell lines were cultured inDMEM high glucose medium containing 10% FBS (Gibco, Rockville, MD, USA), penicillin (100 U/mL) and streptomycin (100 μg/mL) in a 37°C, 5% CO2 incubator. Detection of migrative ability was determined via Transwell assay.
3
Cell Culture Protocols for Various Cell Lines
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A431 (CRL-1555), EA.hy926 (CRL-2922), A498 (HTB-44) and CFPAC-1 (CRL-1918), Jurkat (TIB-152) and RPE-1 (CRL-4000) cells were obtained from the ATCC. They were grown in DMEM (A431, EA.hy926, A498 and CFPAC-1), RPMI medium (Jurkat) or DMEM F12 (RPE-1), supplemented with 10% FBS, and were maintained at 37 °C in a humidified atmosphere of 95% air/5% CO2. Mycoplasma testing was regularly performed.
4
Characterization of Pancreatic Cancer Cell Lines
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The human pancreatic cancer cell lines AsPC1 (CRL-1682), BxPC3 (CRL-1687), CFPAC1 (CRL-1918), MIAPaCa2 (CRL-1420), PANC1 (CRL-1469), and BJ fibroblasts (CRL-2522) were acquired from ATCC, KP4 (JCRB0182) from JCRB, and cultured per supplier’s instructions. HPDE6C7 was provided by Tsao group (U. Toronto) and cultured as previously described31 (link). The spontaneously immortalized human pancreatic stellate cell line hPSC, ONO, and YAM1 were kindly provided by the Evans group (Salk) as previously described11 (link). Normal human fibroblast cells were a kind gift from the Gage group (Salk). MEF cells for Lifr gene knockout characterization were isolated from individual embryos at E13.5, and cultured in DMEM containing 10% FBS for less than five passages. For the low dosage Gem treatment assay, 3 nM gemcitabine or vehicle (PBS) were used.
5
Establishment and Characterization of PDAC Cell Lines
6
Pancreatic Cancer Cell Lines and Mouse Model
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The normal human pancreatic ductal epithelial cells and PC cell lines Panc-1, Bxpc-3, AsPC-1, Capan-1, CFPAC-1, and MIA PaCa-2 were purchased from ATCC (Manassas, VA, USA). These cells were propagated in DMEM added with 10% FBS and 1% antibiotics (penicillin 100 U/mL and streptomycin 100 mg/mL) under 5% CO2 at 37°C. Male BALB/c nude mice (20±2 g, 6–8 weeks) were purchased from SLAC Laboratory Animal Co., Ltd. (Shanghai, China). The mice were housed in a standard laboratory condition (temperature 22°C ±2°C and humidity 50%–60%). All animal experiments were performed in accordance with the guidelines of the Experimental Research Institute of Longhua Hospital Shanghai University of Traditional Chinese Medicine, and were approved by the medical ethics committee of Longhua Hospital Shanghai University of Traditional Chinese Medicine.
7
Silencing UBE2S in Pancreatic Cancer Cells
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Pancreatic cancer cell lines PANC-1 and CFPAC-1 were both obtained from ATCC. PANC-1 cells were cultured in DMEM (Gibco, USA) which was supplemented with 10% fetal bovine serum (FBS). CFPAC-1 cells were cultured in RPMI 1640 (Gibco, USA) which was supplemented with 10% FBS. The cells were grown in 5% CO2-saturated humidity at 37°C and sub-cultured by digesting with trypsin-EDTA.
Pancreatic cell lines PANC-1 and CFPAC-1 were plated in 6-well plates at an appropriate density (40–60% cell density at time of transfection) before transfection with 3 μL UBE2S-RNAi (40 pmol) in a certain amount of serum-free medium, along with the presence of 2 μL lipofectamine 2000, according to the manufacturers’ instructions. The cells were incubated with transfection mixture about 4–6 hrs and that was replaced with fresh medium after transfection. siRNAs for UBE2S and one negative control siRNA were synthesized by Genepharm Technologies (Shanghai, China). The effects of gene silencing were confirmed by Western blotting 48 hrs after transfection.
Pancreatic cell lines PANC-1 and CFPAC-1 were plated in 6-well plates at an appropriate density (40–60% cell density at time of transfection) before transfection with 3 μL UBE2S-RNAi (40 pmol) in a certain amount of serum-free medium, along with the presence of 2 μL lipofectamine 2000, according to the manufacturers’ instructions. The cells were incubated with transfection mixture about 4–6 hrs and that was replaced with fresh medium after transfection. siRNAs for UBE2S and one negative control siRNA were synthesized by Genepharm Technologies (Shanghai, China). The effects of gene silencing were confirmed by Western blotting 48 hrs after transfection.
8
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.
9
Generating Gemcitabine-Resistant Pancreatic Cancer Cells
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The human pancreatic adenocarcinoma cell line CFPAC-1 and AsPC-1 were obtained from ATCC and were cultured in RPMI or DMEM media supplemented with 10% fetal bovine serum. These PDAC cell lines were single-cloned and screened by Western blotting for CD44 expression. To develop gemcitabine-resistant cells, CD44 low single clone cells were transiently exposed to gemcitabine for 16 h once a week with increasing concentrations (50 ng/ml to 1.0 µg/ml) of gemcitabine weekly for more than 2 months. The resulting gemcitabine-resistant cells were referred to as GR and the original gemcitabine-sensitive cells were referred to as GS.
10
Cell Line Authentication and Validation
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The Ca9-22, HEL, NUGC-3, and RPMI8226 cell lines were obtained from the Japanese Collection of Research Bioresources. The 786-O, CFPAC-1, DU145, HCC1599, HCC1806, HCC38, HCT116, HL-60, K-562, MSTO-211H, MV-4-11, NCI-H460, NCI-H2170, and THP-1 cell lines were obtained from the ATCC. The A2780, COLO 792, and DOK cell lines were obtained from the European Collection of Authenticated Cell Cultures. The A549, A673, BHL-89, and MCF-7 cell lines were obtained from DS Pharma Biomedical Co., Ltd. Short-tandem repeat-based DNA profiling was used to reauthenticate cell lines. All cell lines were tested for mycoplasma contamination.
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