Two HCC cell lines, MHCC97-H and MHCC97-L (of the same genetic background but with high and low metastatic potential were previously generated from the parental HCC cell line MHCC97) were obtained from the KeyGEN (Nanjing, China) and kept in Dulbecco Modified Eagle's Medium (Gibco, Grand Island, NY, USA) with 10% heatinactivated fetal bovine serum (Gibco). All cells were fostered in a humidified atmosphere of 5% CO 2 at 37 °C.
Mhcc 97l
Manufactured by Keygen Biotech
Sourced in China
The MHCC-97L is a high-performance cell line used for research purposes. It is a hepatocellular carcinoma cell line derived from a human liver cancer sample. The MHCC-97L cell line is commonly used in scientific research to study liver cancer and related cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Mhcc 97h, by Keygen Biotech (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Hep3b, by Keygen Biotech (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Hepg2, by Keygen Biotech (2 mentions)
Smmc 7721, by Keygen Biotech (2 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Snu423, by Keygen Biotech (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Hek293t cell, by American Type Culture Collection (1 mentions)
Penicillin streptomycin, by Cytiva (1 mentions)
Mhcc97l, by Thermo Fisher Scientific (1 mentions)
Scrambled shrna, by GenePharma (1 mentions)
Mhcc97h, by Thermo Fisher Scientific (1 mentions)
Plc prf 5, by Keygen Biotech (1 mentions)
Thle 2, by Keygen Biotech (1 mentions)
Sk hep 1, by Keygen Biotech (1 mentions)
7 protocols using mhcc 97l
1
Metastatic HCC Cell Line Cultivation
2
Establishment and Maintenance of Liver Cancer Cell Lines
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HCC cell lines (MHCC-97L, MHCC-97H, Hep3B) and immortalized liver cell line (LO2) were procured from KeyGEN BioTECH (JiangSu, China). All cells were maintained in 10% fetal bovine serum (FBS, Gibco Life Technologies, Grand Island, NY, USA) supplemented with high glucose Dulbecco's modified Eagle's medium (DMEM, Invitrogen, Carlsbad, CA, USA) with the addition of 100 U/mL penicillin/streptomycin (Invitrogen, Carlsbad, CA, USA). All cultures were maintained in humidified environments of 5% CO2 at 37°C.
All animal experimental procedures were reviewed and approved by the Southern Medical University Animal Use and Care Committee (Guangdong, China). Female immunodeficient BALB/c-nu/nu mice weighing 20±2 g between 6 to 8 weeks of age were obtained from the Southern Medical University Medical Laboratory Animal Center and reared under specific-pathogen-free (SPF) conditions under a 12 h light/dark cycle. Food and water were provided ad libitum.
All animal experimental procedures were reviewed and approved by the Southern Medical University Animal Use and Care Committee (Guangdong, China). Female immunodeficient BALB/c-nu/nu mice weighing 20±2 g between 6 to 8 weeks of age were obtained from the Southern Medical University Medical Laboratory Animal Center and reared under specific-pathogen-free (SPF) conditions under a 12 h light/dark cycle. Food and water were provided ad libitum.
3
Hepatocellular Carcinoma Tissue Collection
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116 paired pathologically diagnosed HCC samples (including tumor tissues and matched non-tumor liver tissues) were obtained between May 2010 and October 2014 at The Affiliated Changzhou NO.2 People's Hospital of Nanjing Medical University (Changzhou, Jiangsu, China). Written consent approving the usage of tissues in our study were obtained from each patient. The histopathological diagnoses of HCC were conducted by three experienced pathologists independently. This study was approved by the ethnic committee of the Affiliated Changzhou NO.2 People's Hospital of Nanjing Medical University ([2017]KY013-01). Human HCC cell lines (SMMC-7721, Huh7, HepG2, MHCC-97H, MHCC-97L, Hep3B) and the human normal liver L02 cell line were obtained from KeyGen (Nanjing KeyGen Biotech Co., Ltd., Jiangsu, China). All cells used in our study were cultured in Dulbecco's modified Eagle's medium (DMEM, Invitrogen Life Technologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Carlsbad, CA, USA) and 1% streptomycin/penicillin at 37°C in a humidified environment consisting of 95% air and 5% CO2.
4
Hepatocellular Carcinoma Tissue Analysis
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Data were obtained from 108 paired HCC fresh tissues (including tumors and adjacent normal samples) and 12 normal liver tissues (hepatic hemangioma patients) acquired between August 2012 and September 2013 at The First Affiliated Hospital of Nanjing Medical University (Nanjing, Jiangsu, China). Informed consent for tissue analysis was obtained before surgery. The study was approved by the Institutional Ethics Committee of Nanjing Medical University. All research was performed in compliance with government policies and the Helsinki declaration. All experiments were undertaken with the understanding and written consent of each subject. The Huh7, SNU-423, MHCC-97H, MHCC-97L, SMMC-7721, Hep3B, HepG2 human hepatoma cell lines, and the human normal liver L02 cell line were obtained from KeyGen (Nanjing KeyGen Biotech Co., Ltd., Jiangsu, China). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen Life Technologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Carlsbad, CA, USA) at 37 °C in humidified air containing 5% carbon dioxide.
5
Comparative Analysis of HCC Cell Lines
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The MHCC97‐H, MHCC97‐L, and HCCLM3 human HCC cell lines were obtained from KeyGEN (Nanjing, China). Compared with MHCC97‐L, MHCC97‐H has a high metastasis rate.18 Two HCC clones with the same genetic background were established from the MHCC parental hepatocellular carcinoma line. HEK293T cells were obtained from ATCC (Manassas, VA, USA). Cell lines were cultured in 90% DMEM (Gibco, Grand Island, NY, USA) supplemented with antibiotics (13 penicillin/streptomycin 100 U/mL; HyClone Laboratories, Logan, UT, USA) and 10% heat‐inactivated FBS (Gibco). Cells were incubated at 37°C in a humidified atmosphere containing 5% CO2.
6
Metastatic Potential of Hepatocarcinoma Cells
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Human hepatocarcinoma cell lines MHCC97H, MHCC97L and human normal liver cell line L02 were purchased from the KeyGEN Company (Nanjing, China). MHCC97H and MHCC97L cell clones of the same genetic background but with different metastatic potential. The three cell lines were grown in 90% Dulbecco Modified Eagle's Medium (DMEM, Gibco) supplemented with antibiotics (1× penicillin/streptomycin 100 U/ml, Gibco) and 10% heat-inactivated fetal bovine serum (FBS, Gibco). The cells were incubated in a humidified atmosphere of 5% CO2 at 37°C.
shRNA against FUT8, scrambled shRNA, miRNA-34a/miR-26a/miR-455-3p/ normal control (NC) mimics, and miRNA-34a/miR-26a/miR-455-3p/NC inhibitors were chemically synthesized by Shanghai GenePharmaCo., Ltd. (Shanghai, China). MHCC97H and MHCC97L cells were transfected with miRNAs (100 nM, the cell transfection efficiency was 72.3% ~78.5%) or shRNAs (50 nM, the cell transfection efficiency was 76.1%) using Lipofectamine 2000 Transfection Reagent (Invitrogen).
shRNA against FUT8, scrambled shRNA, miRNA-34a/miR-26a/miR-455-3p/ normal control (NC) mimics, and miRNA-34a/miR-26a/miR-455-3p/NC inhibitors were chemically synthesized by Shanghai GenePharmaCo., Ltd. (Shanghai, China). MHCC97H and MHCC97L cells were transfected with miRNAs (100 nM, the cell transfection efficiency was 72.3% ~78.5%) or shRNAs (50 nM, the cell transfection efficiency was 76.1%) using Lipofectamine 2000 Transfection Reagent (Invitrogen).
7
Cell Culture Maintenance Protocol
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Hela, PLC-PRF-5, HepG2, MHCC-97L, MHCC-97H, LO2, THLE2, Hep3B, and SK-Hep-1 cells were purchased from KeyGEN BioTECH and OBIO, Shanghai. Cells are maintained in RPMI 1640 or DMEM medium (Neuronbc) with 10%FBS and 1%PS. All the cell lines were kept at 37 °C under a humidified atmosphere of 5% CO2 in an incubator, trypsinized, and passaged every 2 days. Plasmid DNA was transfected using Lipofectamine 2000 transfection reagent (Invitrogen).
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