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Panc02

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Panc02 is a pancreatic ductal adenocarcinoma cell line derived from a mouse tumor. It is a commonly used in vitro model for pancreatic cancer research.

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Lab products found in correlation

Mia paca 2, by American Type Culture Collection (16 mentions) Panc 1, by American Type Culture Collection (15 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (12 mentions) Bxpc 3, by American Type Culture Collection (11 mentions) Aspc 1, by American Type Culture Collection (6 mentions) Capan 2, by American Type Culture Collection (6 mentions) Hpaf 2, by American Type Culture Collection (6 mentions) Cfpac 1, by American Type Culture Collection (5 mentions) Panc10, by American Type Culture Collection (4 mentions) Panc03, by American Type Culture Collection (4 mentions) Penicillin, by Sartorius (4 mentions) Raw264, by American Type Culture Collection (4 mentions) Penicillin, by Thermo Fisher Scientific (4 mentions) Streptomycin, by Thermo Fisher Scientific (4 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) Capan 1, by American Type Culture Collection (3 mentions) Su 86, by American Type Culture Collection (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) Fetal bovine serum (fbs), by Sartorius (3 mentions) Panc1, by Japanese Collection of Research Bioresources Cell Bank (2 mentions)

43 protocols using panc02

1

Culturing and Viability Assay of PDAC Cell Lines

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The human PDAC cell lines Panc02.03, Panc3.27, Miapaca-2, and the murine PDAC cell lines, Panc02, and KPCP1 were originally procured from ATCC (Manassas, VA). Miapaca-2 was cultured in DMEM medium, and the rest others were cultured in ATCC-recommended RPMI-1640 supplemented with 10% fetal bovine serum and maintained at 5% CO2 at 37 °C. For long-term storage, the cells were frozen in a 5% DMSO containing the respective tissue culture medium in liquid nitrogen. Cell viability assays were carried out using Trypan-blue exclusion method using Beckman Coulter Vi-CELL™ cell viability analyzer and Image analysis [25 ].
Thakur P.C., Miller-Ocuin J.L., Nguyen K., Matsuda R., Singhi A.D., Zeh H.J, & Bahary N. (2018). Inhibition of endoplasmic-reticulum-stress-mediated autophagy enhances the effectiveness of chemotherapeutics on pancreatic cancer. Journal of Translational Medicine, 16, 190.
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2

Syngeneic Tumor Cell Line Maintenance

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The syngeneic mouse tumor cell lines Py8119 breast adenocarcinoma (American Type Culture Collection (ATCC), CRL 3278); MC38 colon carcinoma (Kerafast, ENH204-FP); B16.F10 melanoma (kindly provided by Dr. Dastmalchi, University of Florida); Panc02 pancreatic ductal adenocarcinoma (kindly provided by Dr. Partecke, University of Greifswald); CT26 colon carcinoma (ATCC, CRL-2638); 4T1 breast cancer (ATCC, CRL-2539) were obtained and maintained in their respective media formulations. All the cell culture media were supplemented with 10% fetal bovine serum (FBS), 1% Penicillin-Streptomycin (P/S) antibiotic, and 500 µm of β-mercaptoethanol. Specific medium preparations for each cell line were Py8119 (F-12K base medium); MC38 (DMEM base with 2 mM glutamine, 0.1 mM nonessential amino acids, 1 mM sodium pyruvate, 10 mM HEPES and 50 µg/ml gentamycin sulfate); B16.F10 (DMEM base with 1 mM sodium pyruvate); Panc02 (DMEM base). All cells were maintained at 37°c in 5% CO2- humidified atmosphere. All the cell culture reagents were purchased from Gibco and ATCC.
Sivakumar R., Chan M., Shin J.S., Nishida-Aoki N., Kenerson H.L., Elemento O., Beltran H., Yeung R, & Gujral T.S. (2019). Organotypic tumor slice cultures provide a versatile platform for immuno-oncology and drug discovery. Oncoimmunology, 8(12), e1670019.
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3

Murine Pancreatic Cancer Cell Lines

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The murine pancreatic cancer cell line Panc02 was purchased from the ATCC and cultured in RPMI (Gibco) with 10% FBS, 10 mM L-glutamine, and antibiotics. Murine MT5 (KrasLSL-G12D, Trp53LSL-R270H, Pdx1-cre) pancreatic cells were a kind gift from Dr. Tuveson (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY) and cultured in RPMI (Gibco) with 10% FBS, 10 mM L-glutamine, and antibiotics. KPC-luc cells for orthotopic experiments were derived from KPC mice (KrasLSL-G12D, Trp53−/−, PDX-1-Cre) and transfected with enhanced firefly luciferase as previously described.25 (link) Murine isotype controls (clone 3E5.2H12), anti-IL-6R (clone BP-5875), and anti-PD-L1 blocking Ab were obtained from either Genentech, Inc. (San Francisco, CA) for in vivo studies in the KPC-Brca2 murine model. Murine antibodies to IL-6 (Clone MP5-20F3), PD-L1 (Clone 10F.9G2), or isotype controls (Clones LTF-2 and HRPN) were purchased from BioXcell (West Lebanon, NH) for in vivo studies using the MT-5, Panc02, and KPC-luc cell lines.
Mace T.A., Shakya R., Pitarresi J.R., Swanson B., McQuinn C.W., Loftus S., Nordquist E., Cruz-Monserrate Z., Yu L., Young G., Zhong X., Zimmers T.A., Ostrowski M.C., Ludwig T., Bloomston M., Bekaii-Saab T, & Lesinski G.B. (2016). IL-6 and PD-L1 antibody blockade combination therapy reduces tumor progression in murine models of pancreatic cancer. Gut, 67(2), 320-332.
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4

Characterization of PANC-1 CCK2R knockdown cells

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PANC-1 and Panc02 cell lines (from ATCC) were cultured in DMEM containing 10% FBS. Cells were verified yearly by ATCC STR authentication testing. The characterization of PANC-1 cells stably transfected with a CCK2R shRNA (sh1413) was previously reported [27 (link)].
Abraham T., Armold M., McGovern C., Harms J.F., Darok M.C., Gigliotti C., Adair B., Gray J.L., Kelly D.F., Adair J.H, & Matters G.L. (2024). CCK Receptor Inhibition Reduces Pancreatic Tumor Fibrosis and Promotes Nanoparticle Delivery. Biomedicines, 12(5), 1024.
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5

Multicolor Flow Cytometry for Cell Lines

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Cell Proliferation Dye eFluor 450 ™ (eBioscience) was used for cell proliferation assays, according to the manufacturer’s protocol. Anti-human CD11b-PE (clone ICRF44), anti-human HLA-DR (clone, G46–6)-FITC, anti-human CD14–PerCP-Cy5.5 (clone, mφP9), anti-mouse Gr-1–Alexa Fluor 700 (Biolegend, clone: RB6–8cC5), anti-mouse CD11b–PE-Cy7(clone: M1/70) and Ly-6C-PE (clone AL-21) were obtained from BD Pharmingen. Cell lines used included human head and neck cancer cell lines Cal27, SCC-1, and SCC-25, and human monocytic cell line THP-1. All cell lines were tested free for Mycoplasma. Cell lines were authenticated with GenePrint10 (Promega) through JHH Genetic Resource Core Facility and stored in −70deg C in aliquots when not in use. Cells were cultured with RPMI-1640 medium, with 10% FCS, 10% L-glutamine, 10% Na-pyruvate, 10% Penicillin/Streptomycin and 0.1% beta-ME. All reagents were from Gibco®. When co-cultured with sorted MDSC, the FCS concentration was 1–2%. CD14+ monocytes from healthy non-age matched volunteers were used for controls. Murine melanoma cell line B16, colon-cancer cell line CT26, and pancreatic cancer cell line Panc02 were obtained from ATCC. MOC1 murine head and neck cancer line was obtained from Ravi Uppaluri (Dana Farber).
Zeng Q., Fu J., Korrer M., Gorbounov M., Murray P.J., Pardoll D., Masica D.L, & Kim Y.J. (2018). Caspase-1 from Human Myeloid Derived Suppressor Cells Can Promote T cell–Independent Tumor Proliferation. Cancer immunology research, 6(5), 566-577.
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6

PDAC PDX Cell Lines for IGFBP2 Studies

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Human PDAC PDX cell lines MDA-PATC53 (high endogenous IGFBP2) and MDA-PATC148 (low endogenous IGFBP2) (all KRas G12D) were established as previously reported and cultured in Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 50:50 medium supplemented with 10% fetal bovine serum (FBS) and incubated in a 5% CO2 atmosphere at 37°C [45 (link)]. IGFBP2 stimulation experiments were performed by using recombinant IGFBP2 (Abcam) with cells starved of serum overnight. The mouse PDAC cell line Panc02 was purchased from ATCC (Rockville) and authenticated using short tandem repeat profiling.
Sun L., Zhang X., Song Q., Liu L., Forbes E., Tian W., Zhang Z., Kang Y., Wang H., Fleming J.B., Pasche B.C, & Zhang W. (2020). IGFBP2 promotes tumor progression by inducing alternative polarization of macrophages in pancreatic ductal adenocarcinoma through the STAT3 pathway. Cancer letters, 500, 132-146.
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7

Pancreatic Cancer Cell Lines and CAFs

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The human pancreatic cancer cell lines PK8 and PANC-1 were obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan) and the RIKEN Bio-Resource Center Cell Bank (Ibaraki, Japan). The mouse pancreatic cancer cell line Panc02 is available from ATCC. The murine cancer cell line (Panc02) was provided by T. Moroishi (Department of Cell Signaling and Metabolic Medicine, Faculty of Life Sciences, Kumamoto University). Human CAFs were established from surgically excised pancreatic cancer samples (56 (link)). Mouse CAFs, as described in the literature (57 (link)), were obtained by isolation of fibroblasts from mouse skin and stimulation of the cells with 2 ng/mL recombinant murine TGF-β (R&D Systems) for 6 days. These cells were cultured in RPMI 1640 medium or DMEM containing 10% FBS (normal medium) and incubated at 37°C with 5% CO2.
Kitamura F., Semba T., Yasuda-Yoshihara N., Yamada K., Nishimura A., Yamasaki J., Nagano O., Yasuda T., Yonemura A., Tong Y., Wang H., Akiyama T., Matsumura K., Uemura N., Itoyama R., Bu L., Fu L., Hu X., Wei F., Mima K., Imai K., Hayashi H., Yamashita Y.I., Miyamoto Y., Baba H, & Ishimoto T. (2023). Cancer-associated fibroblasts reuse cancer-derived lactate to maintain a fibrotic and immunosuppressive microenvironment in pancreatic cancer. JCI Insight, 8(20), e163022.
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8

Pancreatic Cancer Cell Line Maintenance

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Standard previously published cell culture methods were used to maintain pancreatic cancer cell lines, including murine Panc02 (NIH repository, DTP/DCTC/NCI[26 (link)]), PanIN 4313 [27 (link)], K8484 and DT8082[28 (link)] as well as human MiaPaca-2, Panc1, and BXPC-3 (ATCC; CRL-1420, CRL-1469, CRL-1687). Luciferase tagged cell lines were generated by infecting the cell lines with a commercial lentiviral titer for firefly-luciferase (Genecopoeia, LPP-FLUC-LV105-025 containing 1x108 TU/ml). Non-infected cells were eliminated after addition of puromycin (10ug/ml, Sigma). Luciferase expression was confirmed for each line through One-Glo analysis (Pierce).
Mendonsa A.M., Chalfant M.C., Gorden L.D, & VanSaun M.N. (2015). Modulation of the Leptin Receptor Mediates Tumor Growth and Migration of Pancreatic Cancer Cells. PLoS ONE, 10(4), e0126686.
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9

Characterization of Melanoma Cell Lines

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B78-D14 [“B78”, obtained from Ralph Reisfeld (Scripps Research Institute) in 2002] melanoma is a poorly immunogenic cell line derived from B78-H1 cells, which were originally derived from B16 melanoma (Becker et al., 1996 (link); Haraguchi et al., 1994 (link); Silagi, 1969 (link)). B78-D14 cells lack melanin, but were transfected with functional GD2/GD3 synthase to express the disialoganglioside GD2 (Becker et al., 1996 (link); Haraguchi et al., 1994 (link)), which is overexpressed on the surface of many human tumors including melanoma (Nazha et al., 2020 (link)). B16-F10 melanoma was obtained from American Type Culture Collection (ATCC) in 2005. The murine pancreatic ductal adenocarcinoma cell line Panc02 was purchased from ATCC. Panc02, B78 and B16 cells were grown in vitro in RPMI-1640 (Mediatech) supplemented with 10% FBS, 2mMol L-glutamine, 100U/ml penicillin, and 100μg/ml streptomycin. Mycoplasma testing via PCR was routinely performed.
Hoefges A., McIlwain S., Erbe A., Mathers N., Xu A., Melby D., Tetreault K., Le T., Kim K., Pinapati R., Garcia B., Patel J., Heck M., Feils A., Tsarovsky N., Hank J., Morris Z., Ong I, & Sondel P. (2023). Antibody landscape of C57BL/6 mice cured of B78 melanoma via immunotherapy. bioRxiv.
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10

Pancreatic Cancer Cell Lines and CAFs

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The human pancreatic cancer cell lines PK8 and PANC-1 were obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan) and the RIKEN Bio-Resource Center Cell Bank (Ibaraki, Japan). The mouse pancreatic cancer cell line Panc02 is available from ATCC. The murine cancer cell line (Panc02) was provided by T. Moroishi (Department of Cell Signaling and Metabolic Medicine, Faculty of Life Sciences, Kumamoto University). Human CAFs were established from surgically excised pancreatic cancer samples (56 (link)). Mouse CAFs, as described in the literature (57 (link)), were obtained by isolation of fibroblasts from mouse skin and stimulation of the cells with 2 ng/mL recombinant murine TGF-β (R&D Systems) for 6 days. These cells were cultured in RPMI 1640 medium or DMEM containing 10% FBS (normal medium) and incubated at 37°C with 5% CO2.
Kitamura F., Semba T., Yasuda-Yoshihara N., Yamada K., Nishimura A., Yamasaki J., Nagano O., Yasuda T., Yonemura A., Tong Y., Wang H., Akiyama T., Matsumura K., Uemura N., Itoyama R., Bu L., Fu L., Hu X., Wei F., Mima K., Imai K., Hayashi H., Yamashita Y.I., Miyamoto Y., Baba H, & Ishimoto T. (2023). Cancer-associated fibroblasts reuse cancer-derived lactate to maintain a fibrotic and immunosuppressive microenvironment in pancreatic cancer. JCI Insight, 8(20), e163022.
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