The human colonic adenocarcinoma epithelial cell line Caco-2 was obtained from Riken Cell Bank (Ibaraki, Japan) and kept in a humidified incubator at 37°C with 5% CO2. Caco-2 cells were maintained in Dulbecco's Modified Eagle Medium (DMEM)-High glucose (Wako, Osaka, Japan) supplemented with 10% heat-inactivated (30 min, 56°C) fetal bovine serum (Biowest, Nuaillé, France), 100 U/mL penicillin, 100 μg/mL streptomycin, and 1% nonessential amino acids (NEAA) (Wako). In this study, Caco-2 cells were used between passages 25 and 40. Cells cultivated to 80% confluence were seeded on 12-well Transwell® inserts (1.12 cm2 polycarbonate membrane with 0.4 μm pore size; Corning Life Sciences, Corning, NY) at a density of 5 × 104 cells per well. The culture medium was changed every 2 days until 20 days later when full polarization of the Caco-2 cell monolayer was achieved. The integrity of the Caco-2 cell monolayer was evaluated by measuring the transepithelial electrical resistance (TER) using a milicell-ERS (Millipore, Billerica, MA). The TER values measured in the experiments were summarized as table form below each figure, and all the figures were expressed as the relative TER value compared to the value before experimental treatment.
Caco 2
Manufactured by Fujifilm
Sourced in Japan
Caco-2 is a laboratory cell line derived from a human colorectal adenocarcinoma. It is commonly used in pharmaceutical research to evaluate the permeability and absorption of drug candidates across the intestinal epithelium.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Fujifilm (2 mentions)
Streptomycin, by Fujifilm (2 mentions)
Ls180, by Fujifilm (2 mentions)
Caco 2, by European Collection of Cell Cultures (2 mentions)
Ls180, by European Collection of Cell Cultures (2 mentions)
Caco 2, by American Type Culture Collection (2 mentions)
Fetal bovine serum (fbs), by Biowest (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Caco 2, by RIKEN Cell Bank (1 mentions)
Non essential amino acids (neaa), by Fujifilm (1 mentions)
Dmem high glucose, by Fujifilm (1 mentions)
Caco 2, by Merck Group (1 mentions)
Transwell insert, by Corning (1 mentions)
Fiveeasyplus fep20 ph meter, by Mettler Toledo (1 mentions)
Fetal bovine serum (fbs), by Cosmo Bio (1 mentions)
Mccoy s 5a, by Thermo Fisher Scientific (1 mentions)
Caco 2, by Sumitomo Pharma (1 mentions)
Hct116, by Thermo Fisher Scientific (1 mentions)
Hct116, by American Type Culture Collection (1 mentions)
7 protocols using caco 2
1
Caco-2 Cell Monolayer for Transepithelial Permeability
2
Culturing Colon Cancer Cell Lines
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The human colon adenocarcinoma cell lines, LS180 and Caco-2, were purchased from European Collection of Cell Cultures (ECACC) collections (KAC, Hyogo, Japan). LS180 cells were cultured in Dulbecco’s modified Eagle medium (DMEM) containing 1,500 mg/L glucose (FUJIFILM Wako Pure Chemical, Osaka, Japan) and Caco-2 cells were cultured in DMEM with 4,500 mg/L glucose. In both the cases, the medium was supplemented with heat-inactivated 10% fetal bovine serum (FBS) (Cosmo Bio, Tokyo, Japan). The cultures were maintained at 37°C in a humidified atmosphere with 5% CO2.
The acidic cell culture medium (pH 6.5) was obtained by the addition of 35–37% HCl solution to DMEM containing 1,500 mg/L glucose supplemented with heat-inactivated 10% FBS by measuring pH with a FiveEasy Plus FEP20 pH Meter (METTLER TOLEDO, Tokyo, Japan). LS180 cells were grown for a sufficient time in normal medium and were subsequently maintained in the acidic cell culture medium (pH 6.5) for a few days as described previously with some modifications [26 (link)–28 (link)].
The acidic cell culture medium (pH 6.5) was obtained by the addition of 35–37% HCl solution to DMEM containing 1,500 mg/L glucose supplemented with heat-inactivated 10% FBS by measuring pH with a FiveEasy Plus FEP20 pH Meter (METTLER TOLEDO, Tokyo, Japan). LS180 cells were grown for a sufficient time in normal medium and were subsequently maintained in the acidic cell culture medium (pH 6.5) for a few days as described previously with some modifications [26 (link)–28 (link)].
3
Cultivation of Cell Lines for Research
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Caco-2 was purchased from DS Pharma Biomedical (Osaka, Japan) and HCT116 was purchased from the American Type Culture Collection. Caco-2 cells were maintained in EMEM (051-07615; Wako) supplemented with 10% fetal bovine serum (FBS; P30-3306; PAN-Biotech GmbH), 5% non-essential amino acids (139-15651; Wako), penicillin, and streptomycin (168-23191; Wako). HCT116 cells were maintained in McCoy's 5A (16600-082; Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS, penicillin, and streptomycin. Human hepatic sinusoidal endothelial cells (HHSECs) were purchased from ScienCell Research Laboratories, Inc. (cat. no. 5000) and maintained with Endothelial Cell Medium (cat. no. 1001; ScienCell Research Laboratories, Inc.) supplemented with 5% FBS and Endothelial Cell Growth Supplement (cat. no. 1052; ScienCell Research Laboratories, Inc.). Cultures were maintained at 37°C in an atmosphere of 95% air and 5% CO2.
4
Culturing Various Human Cell Lines
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HBMECs (human brain microvascular endothelial cells) and HUVECs (human umbilical vein endothelial cells) were purchased from DS Pharma Biomedical (Osaka, Japan). HBMECs and HUVECs were cultured in collagen-coated plates in HuMedia EG-2 (Kurabo, Osaka, Japan). HCF (Human cardiac fibroblasts) were purchased from ScienCell Research Laboratories (Carlsbad, CA). HCF were cultured in RPMI 1640 medium with L-glutamine and 10% FBS. hNHeps™ (Human normal hepatocytes) cells were purchased from Lonza (Lonza, Walkersville, MD). hNHeps™ cells were maintained in hepatocyte culture medium (Lonza). A549 (human lung adenocarcinoma) and HeLa (human epithelial cervical cancer) cells were purchased from Sigma-Aldrich (St. Louis, MO). HepG2 (human hepatocellular carcinoma) cells were provided from Cosmobio (Tokyo, Japan). Caco-2 (human colonic carcinoma) cells were purchased from American Type Culture Collection (Rockville, MD). A549, HeLa, HepG2, and Caco-2 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Wako Pure Chemical Industries, Osaka, Japan) containing 50 units/ml penicillin and 50 μg/ml streptomycin (Sigma-Aldrich, St. Louis, MO), and supplemented with 10% FBS (Wako Pure Chemical Industries). These cells were cultured at 37°C in a humidified atmosphere of 5% CO2.
5
Cell Culture Maintenance Protocols
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HeLa, A549, and THP-1 cell lines were purchased from ATCC, the HaCat cell line was purchased from Cell Lines Service, HUVECs and NHEKs were purchased from PromoCell, and Caco-2 cells were purchased from the Riken Cell Bank. HeLa and A549 cells were maintained in Dulbecco’s modified Eagle’s medium (Nacalai Tesque) supplemented with 10% fetal bovine serum (FBS; Gibco) and 50 μg/ml gentamicin (Nacalai Tesque), and the THP-1 cells were cultured in RPMI 1640 medium (Nacalai Tesque) supplemented with 10% FBS and 50 μg/ml gentamicin. THP-1 cells were differentiated into macrophages by stimulating them with 50 ng/ml phorbol 12-myristate for 72 h. HUVECs were maintained with the endothelial cell growth medium 2 kit (PromoCell) supplemented with 10% FBS and 50 μg/ml gentamicin. NHEKs were cultured with the keratinocyte growth medium 2 kit (PromoCell), and Caco-2 cells were maintained in minimum essential medium (Wako) supplemented with 10% FBS and 50 μg/ml gentamicin. Cells were incubated in a 5% CO2 incubator at 37°C.
6
Culture and Maintenance of Cell Lines
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Cells from the human intestinal epithelial cell line Caco-2, murine macrophage cell line RAW264.7 and murine fibrosarcoma cell line L929 were obtained from American Type culture Collection (ATCC) (Manassas, VA, USA). Human intestinal epithelial cell line, Caco-2, cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM, glutamine, high glucose [Wako]) supplemented with 1% MEM Non-Essential Amino Acids (NEAA [Gibco BRL, Grand Island, NY, USA]), 100 U/ml penicillin, 100 µg/ml streptomycin, and 10% heat-inactivated fetal bovine serum (FBS [Daiichi Kagaku, Tokyo, Japan]). Murine macrophage cell line, RAW264.7, cells were cultured in DMEM (glutamine, low glucose) supplemented with 100 U/ml penicillin, and 100 µg/ml streptomycin, and 10% (v/v) heat-inactivated FBS. Murine fibrosarcoma cell line, L929, cells were cultured in Eagle’s Minimum Essential Medium (MEM [Nissui Pharmaceutical, Tokyo, Japan]) supplemented with 10% FBS, 2 mM L-glutamine, 100 U/ml penicillin and
100 µg/ml streptomycin. The cell cultures were incubated in a humidified 5% CO2 incubator at 37°C.
100 µg/ml streptomycin. The cell cultures were incubated in a humidified 5% CO2 incubator at 37°C.
7
Culture of Colon Adenocarcinoma Cell Lines
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The human colon adenocarcinoma cell lines, LS180 and Caco-2, were purchased from the European Collection of Cell Cultures (ECACC) (EC87021202-F0 and EC86010202-F0; KAC, Hyogo, Japan). LS180 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 1500 mg/L glucose (FUJIFILM Wako Pure Chemical) and Caco-2 cells were cultured in DMEM with 4500 mg/L glucose (FUJIFILM Wako Pure Chemical). In both the cases, the medium was supplemented with heat-inactivated 10% fetal bovine serum (BioWest, Nuaillé, France). The cultures were maintained at 37 °C in a humidified atmosphere with 5% CO2.
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