Fifteen human PC cell lines and one immortalized non-tumorigenic pancreatic epithelial cell line (HPNE) were chosen for this study. All the cell lines were obtained as gifts from Dr Paul J. Chiao’s lab in MD Anderson, Texas University. AsPC-1, AsPC-1 mu [27 (link)], MDA pan28, MDA p28 mu [28 (link)], BxPC-3, Colo357, L3.6pl, MIAPaCa-2, PANC-1, PTAC43, PTAC50, PTAC53, PTAC66, and Capan-1 Cells were grown as a monolayer cell culture in DMEM containing 4.5 mg/ml D-glucose and L-glutamine supplemented with 10% fetal bovine serum. HPNE cells were grown in Medium D with a mixture of M3 medium and DMEM containing one volume of M3 Base F culture medium (InCell Corp., San Antonio, USA), three volumes of glucose-free DMEM, 10% FBS, 5.5 mM glucose, 10 ng/ml EGF, and 50 mg/ml gentamycin. The 293T cell line was grown in complete DMEM. HPNE/Kras/p16shRNA and HPDE/Kras/Her2/p16shRNA/smad4shRNA were pancreatic cancer cell lines (a gift from Dr Paul J. Chiao’s lab, The University of Texas MD Anderson Cancer Center) by transfection of relevant plasmid into HPNE and HPDE cell respectively[29 (link)]. HPDE/Kras/Her2/p16shRNA/smad4shRNA cells were grown in keratinocyte serum-free medium.
M3 base f culture medium
Manufactured by INCELL
Sourced in United States
M3 Base F culture medium is a complete cell culture medium formulation designed to support the growth and maintenance of a variety of cell types. It contains essential nutrients, growth factors, and other components required for cell proliferation and viability.
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Lab products found in correlation
Panc 1, by BNCC (1 mentions)
Hs766t, by BNCC (1 mentions)
Sw1990, by BNCC (1 mentions)
Cfpac 1, by BNCC (1 mentions)
Iscove modified dulbecco medium, by Merck Group (1 mentions)
Cfpac 1, by Merck Group (1 mentions)
Sw1990, by Merck Group (1 mentions)
Leibovitz l 15 medium, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Tideglusib, by Selleck Chemicals (1 mentions)
Hek blue2 cells, by InvivoGen (1 mentions)
Azd1080, by Selleck Chemicals (1 mentions)
Mycoplasma detection kit, by InvivoGen (1 mentions)
Sw620, by American Type Culture Collection (1 mentions)
Du145, by American Type Culture Collection (1 mentions)
Dld 1, by American Type Culture Collection (1 mentions)
Mia paca 2, by American Type Culture Collection (1 mentions)
L3.6pl, by American Type Culture Collection (1 mentions)
4 protocols using m3 base f culture medium
1
Pancreatic Cancer Cell Line Panel
2
Culturing Human Pancreatic Cell Lines
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Human pancreatic ductal cell line (HPNE) and PC cell lines, including PANC-1, SW1990,
HS766T, and CFPAC-1, were purchased from BeNa Culture Collection (Beijing, China) and
cultured in incubator with 100% humidity and 5% CO2 at 37°C. The HPNE cells
were grown in Dulbecco modified Eagle medium (DMEM; Sigma, St Louis, Missouri) containing
1 volume of M3 Base F culture medium (InCell Corp, San Antonio, Texas), 3 volumes of
glucose-free DMEM, 10% fetal bovine serum (FBS; (Invitrogen, Carlsbad, California), 5.5 mM
glucose, 10 ng/mL epidermal growth factor (EGF), and 50 µg/mL gentamycin. The DMEM medium
with 10% FBS was applied to culture PANC-1 and HS766T cells. SW1990 cells were maintained
in Leibovitz L-15 Medium (Sigma). Furthermore, CFPAC-1 cells were cultured in Iscove
Modified Dulbecco Medium (Sigma) adding 10% FBS.
HS766T, and CFPAC-1, were purchased from BeNa Culture Collection (Beijing, China) and
cultured in incubator with 100% humidity and 5% CO2 at 37°C. The HPNE cells
were grown in Dulbecco modified Eagle medium (DMEM; Sigma, St Louis, Missouri) containing
1 volume of M3 Base F culture medium (InCell Corp, San Antonio, Texas), 3 volumes of
glucose-free DMEM, 10% fetal bovine serum (FBS; (Invitrogen, Carlsbad, California), 5.5 mM
glucose, 10 ng/mL epidermal growth factor (EGF), and 50 µg/mL gentamycin. The DMEM medium
with 10% FBS was applied to culture PANC-1 and HS766T cells. SW1990 cells were maintained
in Leibovitz L-15 Medium (Sigma). Furthermore, CFPAC-1 cells were cultured in Iscove
Modified Dulbecco Medium (Sigma) adding 10% FBS.
3
HPNE, 293T, and HPDE/K-ras Cell Culture Protocol
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The previously described [13] (link) HPNE cell line was obtained from Dr. James W. Freeman at The University of Texas Health Science Center at San Antonio (Texas). HPNE cells were grown in Medium D with a mixture of M3 medium and DMEM containing one volume of M3 Base F culture medium (InCell Corp., San Antonio, USA), three volumes of glucose-free DMEM, 10% FBS, 5.5 mM glucose, 10 ng/ml EGF, and 50 µg/ml gentamycin. The 293T cell line was grown in complete DMEM. HPDE/K-ras cells were grown in keratinocyte serum-free medium.
4
Comprehensive Cancer Cell Line Panel
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Human lung (Calu-6, A549, H460, H661, H2126, H322, H1299, H522, PC9, and H4006), colon (SW620, DLD-1, and HCT-8), pancreatic (MiaPaCa2 and L3.6pl), and prostate (DU145) cancer cell lines and HEK293 cells were obtained from the American Type Culture Collection and cultured in Dulbecco’s modified Eagle’s medium (DMEM) or RPMI-1640 medium. Normal/immortalized T80 cells (J. Liu and R. Bast, MD Anderson Cancer Center) were cultured in Medium 199/MCDB 105. hTERT-immortalized HPNE cells (Channing Der, University of North Carolina) were grown in medium D (mixture of M3 medium and DMEM) containing one volume of M3 Base F culture medium (InCell Corp.), three volumes of glucose-free DMEM, 5.5 mM glucose, 10 ng/mL EGF, and 50 µg/mL gentamycin. All media were supplemented with 10% heat-inactivated fetal bovine serum, 10 U/mL penicillin, and 10 µg/mL streptomycin. SB was synthesized in-house as described previously30 (link) and dissolved in dimethyl sulfoxide (DMSO; Sigma-Aldrich). Tideglusib, AZD1080, and BIO were purchased from SelleckChem. The Published Kinase Inhibitor Set 1 (PKIS1) of 304 compounds was received from GlaxoSmithKline (GSK). All cell lines were mycoplasma-free, monitored regularly with HEK-blue2 cells and mycoplasma detection kit from invivogen (cat# rep-pt1).
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