Human CRC cell lines SW480, SW620, HT29, Colo205, Caco-2 and HCT116 were purchased from Shanghai Zhong Qiao Xin Zhou Biotechnology Co.,Ltd (Shanghai, China). Caco-2, SW620 and Colo205 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), HCT116 cells were cultured in DMEM medium supplemented with 10% FBS, HT29 were cultured in McCoy’s 5A medium supplemented with 10% FBS, SW480 were cultured in Leibovitzs L-15 medium supplemented with 10% FBS. All types of culture mediums were purchased from Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd. All CRC cells were maintained in an incubator under the condition of 5% CO2 at 37°C.
Caco 2
Manufactured by Shanghai Zhong Qiao Xin Zhou Biotechnology
Sourced in China
Caco-2 is a human epithelial colorectal adenocarcinoma cell line that is commonly used in in vitro studies to evaluate drug absorption and permeability. It is a well-established model for assessing the intestinal permeability of pharmaceutical compounds.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Colo 205, by Shanghai Zhong Qiao Xin Zhou Biotechnology (2 mentions)
Sw620, by Shanghai Zhong Qiao Xin Zhou Biotechnology (2 mentions)
Hct116, by Shanghai Zhong Qiao Xin Zhou Biotechnology (2 mentions)
Sw480, by Shanghai Zhong Qiao Xin Zhou Biotechnology (2 mentions)
Ht 29, by Shanghai Zhong Qiao Xin Zhou Biotechnology (2 mentions)
Sw620, by Thermo Fisher Scientific (1 mentions)
Sw480, by Thermo Fisher Scientific (1 mentions)
Ly294002, by Cell Signaling Technology (1 mentions)
S4441, by Merck Group (1 mentions)
Cleaved poly adp ribose polymerase parp, by Cell Signaling Technology (1 mentions)
Caco 2, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
D2650, by Merck Group (1 mentions)
Bcl2 associated x (bax), by Santa Cruz Biotechnology (1 mentions)
Bcl 2, by Santa Cruz Biotechnology (1 mentions)
Dmem medium, by BasalMedia (1 mentions)
4 protocols using caco 2
1
Culture and Maintenance of CRC Cell Lines
2
Culturing of Colorectal Cancer Cell Lines
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CC cells, including SW480, Colo-205, SW620, HCT15, DLD-1, HCT116, Caco-2, RKO and LOVO were bought from Shanghai Zhong Qiao Xin Zhou Biotechnology. SW480 and SW620 were cultured in a complete DMEM medium supplemented with 10% FBS (Gibco Grand Island, NY, USA). Colo-205, HCT15 and DLD-1 cells in a complete RPMI-1640 medium supplemented with 10% FBS. Caco-2 and RKO cells in a complete EMEM medium supplemented with 10% FBS. HCT116 cells in a complete Mccoy’s 5A medium supplemented with 10% FBS. LOVO cells in a complete F-12K medium supplemented with 10% FBS. All the experimental procedures described below were repeated thrice unless described otherwise.
3
Exploring Colorectal Cancer Cell Lines
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The human colorectal adenocarcinoma cancer cell lines Caco-2 and CX-1 were purchased from Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd. Caco-2 and CX-1 cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) and RPMI 1640 (Gibco, Thermo Fisher Scientific), respectively, supplemented with 10% fetal bovine serum (BI, Israel) and 1% penicillin/streptomycin (PEN/STREP 100×, MILLIPORE, USA). Cells were cultured at 37°C in a 5% CO2 and 95% air humidified incubator. Stocks of SAL (S4526, Sigma-Aldrich, USA) and SFN (S4441, Sigma-Aldrich, USA) were prepared via dissolution in dimethyl sulfoxide (DMSO, D2650, Sigma-Aldrich, USA) at concentrations of 50 mM and 100 mM, respectively, and the solutions were stored at −20°C. The final concentration of DMSO was less than 0.1%. Antibodies against PI3K (1:1000 dilution), Akt (1:1000 dilution), phosphorylated Akt (Ser473) (1:1000 dilution), p53 (1:1000 dilution), cleaved poly ADP-ribose polymerase (PARP) (1:1000 dilution), and β-actin (1:1000 dilution) as well as the PI3K inhibitor LY294002 were purchased from Cell Signaling Technology. Antibodies against Bcl-2 (1:100 dilution) and Bax (1:100 dilution) were obtained from Santa Cruz Biotechnology.
4
Colorectal Cancer Cell Line Manipulation
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The human colorectal adenocarcinoma cell lines (Caco-2 and HT-29) were purchased from Shanghai Zhongqiao Xinzhou Biotechnology Co., Ltd. and maintained in DMEM medium (BasalMedia, Shanghai, China) supplemented with 10% FBS (Thermo Fisher Scientific, Waltham, MA, United States) and incubated at 5% CO2 at 37°C.
The MMP9 sequence was synthesized by General Biol and then inserted into the pCDH-CMV-MCS-EF1-copGFP-T2A-Puro lentiviral expression vector plasmids (General Biol, Anhui, China), called MMP9 OE plasmids. In addition, lentivirus-containing shRNA targeting MMP9 (MMP9 shRNA1: 5′-GGAATACCTGTACCGCTATGG-3′, MMP9 shRNA2: 5′- GCAGACATCGTCATCCAGTTTC-3′ and MMP9 shRNA3: 5′-GCTTAGATCATTCCTCAGTGC-3′) were synthesized by GenePharma (Shanghai, China). After that, 293T cells were transfected with indicated lentiviral plasmids, packaging plasmids (pAX2) and envelope plasmid (pMD2. G) for 72 h. Next, virus-containing supernatants was filtered through a filter membrane (0.22 μm pore size), and then transduced into Caco-2 and HT-29 cells. Later on, the infected cells were selected by puromycin.
The MMP9 sequence was synthesized by General Biol and then inserted into the pCDH-CMV-MCS-EF1-copGFP-T2A-Puro lentiviral expression vector plasmids (General Biol, Anhui, China), called MMP9 OE plasmids. In addition, lentivirus-containing shRNA targeting MMP9 (MMP9 shRNA1: 5′-GGAATACCTGTACCGCTATGG-3′, MMP9 shRNA2: 5′- GCAGACATCGTCATCCAGTTTC-3′ and MMP9 shRNA3: 5′-GCTTAGATCATTCCTCAGTGC-3′) were synthesized by GenePharma (Shanghai, China). After that, 293T cells were transfected with indicated lentiviral plasmids, packaging plasmids (pAX2) and envelope plasmid (pMD2. G) for 72 h. Next, virus-containing supernatants was filtered through a filter membrane (0.22 μm pore size), and then transduced into Caco-2 and HT-29 cells. Later on, the infected cells were selected by puromycin.
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