Murine colon carcinoma CT26, human bladder carcinoma J82, T24, SW1710, and UM-UC-3, breast cancer MCF-7 and T47D, colorectal cancer DLD-1, HCT15, HT29, Lovo, and HCT116, hepatoma HepG2, non-small cell lung cancer NCI-H460, ovary cancer Caov-3, prostate cancer LNCap clone FGC, and renal carcinoma Caki-1, as well as normal HUVEC, intestinal epithelial FHs74Int, and lung fibroblast MRC-5 cell lines were obtained from the National Collection of Authenticated Cell Cultures. Murine colorectal adenocarcinoma MC38 cell lines were purchased from MingzhouBio, Ningbo, Zhejiang, China. The cells were maintained in the logarithmic phase of growth in DMEM, MEM, McCoy’5a, or RPMI medium supplemented with 10% heat-inactivated FBS, 100 IU/mL penicillin, and 100 μg/mL streptomycin in an incubator at 37 °C with a humidified atmosphere of 5% CO2. Cell line identity was validated through short tandem repeat profiling, and routine mycoplasma testing was negative for contamination.
Hct15
Manufactured by National Collection of Authenticated Cell Cultures
Sourced in China
The HCT15 is a laboratory equipment used for cell culture. It is a well-established cell line derived from human colorectal adenocarcinoma. The HCT15 cell line is a widely used tool in cancer research and drug development.
Automatically generated - may contain errors
Lab products found in correlation
Hct116, by National Collection of Authenticated Cell Cultures (9 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Dld 1, by National Collection of Authenticated Cell Cultures (3 mentions)
Sw620, by National Collection of Authenticated Cell Cultures (3 mentions)
Sw480, by National Collection of Authenticated Cell Cultures (3 mentions)
Hepg2, by National Collection of Authenticated Cell Cultures (2 mentions)
Sw1710, by National Collection of Authenticated Cell Cultures (2 mentions)
Umuc3, by National Collection of Authenticated Cell Cultures (2 mentions)
Mcf 7 and t47d, by National Collection of Authenticated Cell Cultures (2 mentions)
Nci h460, by National Collection of Authenticated Cell Cultures (2 mentions)
Caov3, by National Collection of Authenticated Cell Cultures (2 mentions)
Normal huvec, by National Collection of Authenticated Cell Cultures (2 mentions)
Fhs74int, by National Collection of Authenticated Cell Cultures (2 mentions)
Lncap clone fgc, by National Collection of Authenticated Cell Cultures (2 mentions)
Mrc 5, by National Collection of Authenticated Cell Cultures (2 mentions)
Caki 1, by National Collection of Authenticated Cell Cultures (2 mentions)
Hct15, by Thermo Fisher Scientific (2 mentions)
Antibiotic solution, by ScienCell (1 mentions)
Endothelial cell growth supplements, by ScienCell (1 mentions)
Humidified incubator, by Thermo Fisher Scientific (1 mentions)
11 protocols using hct15
1
Cell Line Maintenance and Characterization
2
Cell Line Maintenance and Characterization
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Murine colon carcinoma CT26, human bladder carcinoma J82, T24, SW1710, and UM-UC-3, breast cancer MCF-7 and T47D, colorectal cancer DLD-1, HCT15, HT29, Lovo, and HCT116, hepatoma HepG2, non-small cell lung cancer NCI-H460, ovary cancer Caov-3, prostate cancer LNCap clone FGC, and renal carcinoma Caki-1, as well as normal HUVEC, intestinal epithelial FHs74Int, and lung fibroblast MRC-5 cell lines were obtained from the National Collection of Authenticated Cell Cultures. Murine colorectal adenocarcinoma MC38 cell lines were purchased from MingzhouBio, Ningbo, Zhejiang, China. The cells were maintained in the logarithmic phase of growth in DMEM, MEM, McCoy’5a, or RPMI medium supplemented with 10% heat-inactivated FBS, 100 IU/mL penicillin, and 100 μg/mL streptomycin in an incubator at 37 °C with a humidified atmosphere of 5% CO2. Cell line identity was validated through short tandem repeat profiling, and routine mycoplasma testing was negative for contamination.
3
Cell Culture Procedure for HCT116, RKO, and HCT15
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HCT116, RKO and HCT15 cell lines were purchased from the Cell Bank of Chinese Academy of Sciences (Shanghai, China). The three cell lines were respectively grown in McCoy’s 5 A medium, minimun essential medium or RPMI 1640 medium containing 10% fetal bovine serum. Cell passage was carried out in a clean platform. After digestion and centrifugation, the cells were resuspended with complete medium, planted on cell culture dishes or plates and cultured in a humidified atmosphere containing 5% CO2 at 37 °C.
4
Cell Culture Conditions for CRC and HUVEC
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Two human CRC cell lines HCT116 and HCT-15 were obtained from the Cell Bank of Chinese Academy of Sciences (Shanghai, China), and were cultured in DMEM medium supplemented with 10% FBS, 100 µg/ml of penicillin, and 100 mg/ml of streptomycin. The Human Umbilical Vascular Endothelial Cells (HUVECs) were purchased from Kelei Corporation (Shanghai China) and cultured on plates coated with 30 μg/ml vitrogen (Collagen Biomaterials, Palo Alto, CA) in ECM medium supplemented with 10% fetal bovine serum, endothelial cell growth supplements and antibiotic solution (Sciencell Research Laboratories, USA). All cells had been validated by short tandem repeat (STR) profiling. Cells were cultured at 37℃ with 5% CO2 in a humidified incubator (Thermo, Waltham, MA) [34] (link).
5
Cell Culture Conditions for Colon Cancer Research
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Normal human colon mucosal epithelial cell line NCM460 and colon cancer cell lines HT-29, SW620, HCT-15, and HCT116 were purchased from Cell Bank of Chinese Academy of Sciences (Shanghai, China). NCM460 cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) (GIBCO). HT-29 and HCT116 cells were grown in McCoy’s 5a medium (Sigma). SW620 cells were grown in L-15 medium (GIBCO). HCT-15 cells were grown in RPMI-1640 medium (GIBCO). All cells were cultured in the above described medium complemented with 10% fetal bovine serum (GIBCO) in a moist air with 5% CO2 at 37°C. Where indicated, cells were treated with 1 μM GDC-0994 (Selleck) for the indicated time.
6
Establishing Acidic CRC Cell Lines
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Human colorectal cancer (CRC) cell lines HCT15 and HCT116 were obtained from the Cell Bank of Chinese Academy of Sciences (Shanghai, China). The cells were cultured in RPMI-1640 (pH 7.4), supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin. Cells were maintained in a 5% CO2/95% air incubator at 37°C. Acidic medium was prepared by adjusting the pH to 6.5 with 25 mmol/L each of PIPES and HEPES. CRC-AA cells (HCT15-AA and HCT116-AA) were obtained by continuously culturing and passing the CRC cells in acidic medium for at least three months.
7
Culturing Human Colon Cancer Cells
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8
Culturing and Transfecting CRC Cell Lines
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CRC cell lines including RKO, HCT116, SW480, SW620, HCT15, DLD1, HT29 and Caco2 were obtained from Cell Bank of the Chinese Academy of Sciences (Shanghai, China), and maintained as previously described [16 (link)]. All the cells were cultured in RPMI 1640 (Hyclone; Logan, Utah, USA) with 10% fetal bovine serum (FBS) (Gibco-BRL, Invitrogen; Paisley, UK) at 37 °C with a humidity of 5% CO2. Plasmids including pcDNA3-LASP1, and pcDNA3-CCT8 were constructed in our lab. All siRNA oligos including LASP1 and CCT8 specific siRNAs were purchased from GenePharma (Shanghai, China). The siRNA oligos used are shown in Supplementary Table S1 . CRC cells at exponential growth phase were plated into 6-well plates for 24 h at a density of 0.5 × 105 cells/mL, and transfected with 1 mg of siRNA or 4 µg cDNA in reduced serum medium (OPTI-MEM-I; Invitrogen) according to the manufacturer’s protocol.
9
Erythroferrone Expression Quantification
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HCoEPIC (Item No. CP-H122) was purchased from Tongpai Biotechnology Co., Ltd. (Shanghai, China). SW480, SW620, HCT116, HCT15, HT29, and CT26 were purchased from the Cell Bank of the Chinese Academy of Sciences. Cells were added with the corresponding medium according to the instructions and placed in a cell culture incubator at 37 °C with 5% CO2. After RNA extraction from the cells, RNA reverse transcription was performed using the PrimeScript™ RT kit from Takara, Japan, and ERFE mRNA expression was detected by SYBR® premix ExTaq™ II (Takara). RT-qPCR was performed using a fluorescence quantitative PCR instrument-9 LightCycler 480II (ROCHE). RT-qPCR primers for ERFE and β-actin were as follows: ERFE forward, 5′-ATG-GGG-CTG-GAG-AAC-AGC-3′, reverse, 5′-TGG-CAT-TGT-CCA-AGA-AGA-CA-3′; and β-actin forward, 5′-AGGCTCTTTTCCAGCCTTCC -3′, reverse, 5′-CTGTCAGCAATGCCAGGGGTA-3′.
10
Authenticated Cell Lines for Colorectal Cancer Research
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Human colorectal cancer cell lines HCT116, RKO, HCT15, and HT29 were obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). The human peritoneal mesothelial cell line HMrSV5 (SV5) was provided by Prof. Huimian Xu (Department of Surgical Oncology and General Surgery, The First Hospital of China Medical University). All cell lines were cultured with RPMI-1640 containing 10% fetal bovine serum (FBS, Gibco, Gaithersburg, MD, USA) and 1% penicillin/streptomycin. The short tandem repeats (STR) profile was used to authenticate all the cell lines. All cells were cultured for no longer than 2 months and negatively tested for mycoplasma contamination.
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