The cell lines SMMC-7721 and A549 were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and were cultured in complete DMEM (Wisent Inc., St-Bruno, QC, Canada) supplemented with 10% fetal bovine serum (Wisent Inc.) at 37°C in a humidified incubator with 5% CO2.
Complete dmem
Manufactured by Wisent
Sourced in China, United States
Complete DMEM is a cell culture medium formulated to support the growth and maintenance of a wide variety of cell types. It provides essential nutrients, vitamins, and other components required for cell proliferation and survival.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Wisent (4 mentions)
Smmc 7721, by National Collection of Authenticated Cell Cultures (1 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Recombinant il 4, by Thermo Fisher Scientific (1 mentions)
Myo inositol, by Merck Group (1 mentions)
Rsm 932a, by Cayman Chemical (1 mentions)
Hemicholinium 3, by Merck Group (1 mentions)
L methionine, by Merck Group (1 mentions)
Hcc lm3, by Wisent (1 mentions)
Smmc 7721, by Wisent (1 mentions)
Complete rpmi 1640 medium, by Wisent (1 mentions)
4 protocols using complete dmem
1
Cell Line Culture Procedure
2
Silencing and Overexpressing DEPDC1B in Human Pancreatic Cell Lines
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All human PC cell lines and immortalized human pancreatic ductal cell lines were obtained from the American Type Culture Collection (Rockville, MD, USA) and were cultured in complete DMEM (Wisent, Inc., St-Bruno, QC, Canada) containing 10% fetal bovine serum (Wisent, Inc.) at 37°C in a 5% CO2 humidified incubator.
To create DEPDC1B-silenced cell lines, PANC-1 cells and CFPAC-1 cells were transfected with DEPDC1B-siRNA and negative control-siRNA (synthesized by Shanghai GenePharma Co., Ltd., Shanghai, China) using Lipofectamine 3000 reagent (Invitrogen, Carlsbad, California, USA) according to the instructions. Following transfection for 48–72 h, the cells were collected for subsequent experiments. To establish stable DEPDC1B-overexpressing cell lines, SW1990 cells and BXPC3 cells were transfected with LV-DEPDC1B and LVCON319 (designed by Shanghai GenePharma Co., Ltd., Shanghai, China) according to the manufacturer’s instructions. The knockdown and overexpression efficiencies were confirmed by Western blotting. All sequences used were as follows: DEPDC1B#1-siRNA, GCAAGCAGGGAGUUGUUAUTT; DEPDC1B#2-siRNA, GCACUUCACAAGAGAACAUTT; NC-siRNA, GUACUGGGUUUGUUACAGAUU.
To create DEPDC1B-silenced cell lines, PANC-1 cells and CFPAC-1 cells were transfected with DEPDC1B-siRNA and negative control-siRNA (synthesized by Shanghai GenePharma Co., Ltd., Shanghai, China) using Lipofectamine 3000 reagent (Invitrogen, Carlsbad, California, USA) according to the instructions. Following transfection for 48–72 h, the cells were collected for subsequent experiments. To establish stable DEPDC1B-overexpressing cell lines, SW1990 cells and BXPC3 cells were transfected with LV-DEPDC1B and LVCON319 (designed by Shanghai GenePharma Co., Ltd., Shanghai, China) according to the manufacturer’s instructions. The knockdown and overexpression efficiencies were confirmed by Western blotting. All sequences used were as follows: DEPDC1B#1-siRNA, GCAAGCAGGGAGUUGUUAUTT; DEPDC1B#2-siRNA, GCACUUCACAAGAGAACAUTT; NC-siRNA, GUACUGGGUUUGUUACAGAUU.
3
Isolation and Polarization of Bone Marrow-Derived Macrophages
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BMDM were isolated and cultured as previously described [80 (link), 81 (link)]. Briefly, bone marrow cells were obtained from the femur and tibia by centrifugation [4 (link)]. Cells were differentiated into macrophages using 15–20% L929-conditioned media in complete DMEM (Wisent) containing 10% FBS (Wisent), 1% penicillin/streptomycin. Cells were plated into 15 cm dishes and allowed to differentiate for 6–8 days. Cells were lifted by gentle scraping in 10mM EDTA in PBS, counted, and seeded into culture plates for experiments at 1x106/mL. Cells were treated for 24 h with DMSO as vehicle or inhibitors: hemicholinium-3 (Sigma-Aldrich), RSM-932a (Cayman Chemicals). Macrophages were polarized with 20 ng/mL recombinant IL-4 (Peprotech or Roche) or 100 ng/mL LPS (E. coli:B4, Sigma-Aldrich). Choline-free DMEM was formulated by preparing nutrient-deficient DMEM (US Biologicals, D9809) according to manufacturer’s instructions and supplementing with sodium pyruvate (Gibco), myo-inositol, L-methionine, and calcium D-pantothenate (Sigma-Aldrich).
4
Culturing Diverse Liver Cancer Cell Lines
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The LO2 (human embryonic liver) cell line and the Huh7 (human HCC) cell line were purchased from the Cell Bank of Xiangya Central Laboratory, Central South University. Three liver cancer cell lines, HCCLM3, MHCC‐97 L, and MHCC‐97H, were gifts from the Liver Cancer Institute, Zhongshan Hospital, Fudan University. SMMC‐7721, HepG2, Bel‐7402, Hep3B, PLC, and Li‐7 cell lines were obtained from the cell bank of the Chinese Academy of Sciences. HCCLM3, MHCC‐97 L and MHCC‐97H, HepG2, Hep3B, SMMC‐7721, PLC, Huh7, and Li‐7 cells were cultured in complete DMEM (Wisent, Inc.). LO2 and Bel‐7402 cells were cultured in complete RPMI‐1640 medium (Wisent, Inc.). All cells were supplemented with 10% fetal bovine serum (Wisent, Inc.) at 37°C in a humidified incubator with 5% CO2.
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