Thyroid cancer cell line BHT101 and BCPAP was purchased from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). BHT101 cell was grown in DMEM medium (Gibco) containing 20% heat‐inactivated FCS (Gibco) and glutamine; BCPAP cells was maintained in RPMI 1640 medium containing 10% fetal bovine serum (Gibco) and 2 mmol/L l ‐glutamine (Invitrogen, Carlsbad, CA) at 37°C with 5% CO2. The LINC00704 and negative control siRNAs (Invitrogen, Carlsbad, CA) were transfected into BHT101 and BCPAP cells using RNAiMAX (Invitrogen) according to the manufacturer's protocol. Forty‐eight hours after transfection, the cells were harvested for further experiment. The LINC00704 siRNA sequences are siRNA 1#, 5′‐GCUUGCUCUCACAGCCAUUTT‐3′ siRNA 2#, 5′‐GGAACCUCCUUUCUUACAUTT‐3′.
Bht 101
Manufactured by Thermo Fisher Scientific
Sourced in United States
The BHT-101 is a laboratory equipment manufactured by Thermo Fisher Scientific. It is a benchtop hot plate that provides consistent and precise temperature control for a variety of laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions)
Nthy ori 3 1, by Thermo Fisher Scientific (4 mentions)
Bcpap, by Thermo Fisher Scientific (3 mentions)
Bcpap, by National Collection of Authenticated Cell Cultures (3 mentions)
Dmem medium, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Bht 101, by National Collection of Authenticated Cell Cultures (2 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
Kmh 2, by Thermo Fisher Scientific (2 mentions)
Cal 62, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Rnaimax, by Thermo Fisher Scientific (1 mentions)
Nthy ori 3 1, by Jennio Biotech (1 mentions)
Ftc 133, by Procell (1 mentions)
Tpc 1, by BNCC (1 mentions)
Nthy ori 3 1, by BNCC (1 mentions)
Tpc 1, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
8 protocols using bht 101
1
Thyroid Cancer Cell Line Manipulation
2
Culturing Thyroid, DTC, and Retinal Cells
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The human normal thyroid cell Nthy-ori 3-1was purchased from Jennio biotech (Guangzhou, China). DTC cell line FTC-133 and human retinal microvascular endothelial cells (RCECs) were obtained from Procell life science and technology (Wuhan, China). DTC cell line B-CPAP, human BHT-101 cells and ACT cells were obtained from Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). The Nthy-ori 3 − 1, BHT101, B-CPAP, BHT-101 and ACT cell line were maintained in RPMI 1640 medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS) (Sigma, St. Louis, MO, USA), 100 U/ml of penicillin G and 100 μg/ml of streptomycin. RCECs were cultured in Dulbecco’s Modified Eagle’s medium (DMEM) (Gibco Life Technologies, Rockville, MD, USA) supplemented with 10% FBS, 100 U/ml of penicillin G and 100 μg/ml of streptomycin. The cultures were maintained in a humidified atmosphere of 5% CO2 and 95% air at 37 ˚C.
3
In Vitro Culture of Thyroid Cell Lines
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Human thyroid normal cell line Nthy-ori 3-1 and thyroid cancer cell lines TPC-1 and BHT-101 were obtained from BeNa Culture Collection (Beijing, China). RPMI-1640 medium (Gibco) added with 10% fetal bovine serum (FBS; Gibco) and 10% penicillin (100 units/mL)–streptomycin (100 μg/mL) mixed solution was used for the cultivation of Nthy-ori 3-1 and TPC-1 cells. BHT-101 cells were cultivated with DMEM (Gibco) added with 20% FBS (Gibco), 100 units/mL penicillin and 100 μg/mL streptomycin. Cells were grown in a 37°C, 5% CO2 humidified incubator.
4
NRARP Knockdown in Thyroid Cancer Cells
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Human ATC cell lines BHT-101[10 (link)] and 8305C[11 (link)] were purchased from China Center for Type Culture Collection (Shanghai, China). Normal human thyroid follicular cell line Nthy-ori 3-1 was purchased from the European Collection of Cell Cultures (Salisbury, UK). Nthy-ori 3-1 was grown at 37°C in complete Roswell Park Memorial Institute 1640 medium (GIBCO, USA), while BHT-101 and 8305C were maintained at 37°C in Dulbecco's modified Eagle medium (GIBCO, USA). All media were supplemented with 10% fetal bovine serum (Sigma, USA) and 1% penicillin/streptomycin (GIBCO, USA) in 5% CO2 incubator. Subcultures were maintained at 80% confluence and passaged by 0.25% trypsin (GIBCO, USA).
To produce cells deficient in NRARP, Lentivirus carrying NRARP-shRNA (Lenti-NRARP-shRNA) (Santa Cruz) and Lenti-CON (Santa Cruz) were transfected into cells. In brief, BHT-101 and 8305C were cultured in 12-well plates for 24 h, then 5 μg/ml Lenti-NRARP-shRNA or Lenti-CON was added into the wells. After 24 h with infection, cells were cultured in fresh medium overnight. To obtain stable-transfected cells lines, cells were subcultured with 5 ug/ml puromycin for 2 weeks.
To produce cells deficient in NRARP, Lentivirus carrying NRARP-shRNA (Lenti-NRARP-shRNA) (Santa Cruz) and Lenti-CON (Santa Cruz) were transfected into cells. In brief, BHT-101 and 8305C were cultured in 12-well plates for 24 h, then 5 μg/ml Lenti-NRARP-shRNA or Lenti-CON was added into the wells. After 24 h with infection, cells were cultured in fresh medium overnight. To obtain stable-transfected cells lines, cells were subcultured with 5 ug/ml puromycin for 2 weeks.
5
Culture of Thyroid Cancer Cell Lines
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Human anaplastic thyroid carcinoma cell lines BHT-101 and KMH-2 were purchased from Cell Bank of the Chinese Academy of Sciences (Shanghai, China).[14 (link)15 (link)] Normal human thyroid follicular cell line Nthy-ori 3-1 was purchased from the Institute of European Collection of Cell Cultures (ECACC, UK). Nthy-ori 3-1 was grown at 37°C in complete RPMI 1640 (GIBCO Inc., MD, USA). BHT-101 and KMH-2 cells were maintained at 37°C in Dulbecco's modified Eagle's medium (GIBCO Inc.). All media were supplemented with 10% fetal bovine serum (Hyclone, UT, USA), 1 mmol/L nonessential amino acid and 1% penicillin/streptomycin (Sigma-Aldrich Co., MO, USA) in 5% CO2 incubator. Subcultures were maintained at 80% confluence and passaged by 0.25% Trypsin (GIBCO Inc.).
6
Thyroid Cancer Cell Lines and Tissues
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Human thyroid cancer cell lines CAL-62, KHM-5M, BHT-101, B-CPAP, and normal thyroid cell lines Nthy-ori-3-1 were purchased from the China Center for Type Culture Collection (CCTCC, China). The CAL-62 and BHT-101 cells were cultured in DMEM medium supplemented with 10% and 20% fetal bovine serum (Gibco, USA) at 37°C in 5% CO2, respectively. KHM-5M, B-CPAP and Nthy-ori-3-1 were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (Gibco, USA) at 37°C in 5% CO2. A total 15 of DTC patients who underwent radical thyroidectomy in Ruijin Hospital of Shanghai Jiaotong University Medical College and were confirmed as DTC by postoperative histopathological examination were included in this study. All samples were obtained with the patients’ informed consent, and the samples were histologically confirmed by at least 2 pathologists independently in a double-blinded fashion.
7
Cell Culture Conditions for CAL-62 and BHT-101
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CAL-62 and BHT-101 cell lines were purchased from FuHeng Cell Center (Shanghai, China) and cultured in DMEM medium (KeyGen BioTECH, Jiangsu, China) supplemented with 10% fetal bovine serum (FBS) (Gibco, NY, USA) for CAL-62 and 20% FBS for BHT-101. A total of 4–6 × 104 cells/mL cells were plated into culture dishes (Corning, NY, USA) and incubated in atmosphere of 5% CO2 at 37 °C for 12–24 h before the experiments were performed. For immunofluorescent labeling, several cell-bearing coverslips were prepared under similar conditions by using coverslip preparation dishes (JET Bio-Filtration, Guangzhou, China) for multiple experiments.
8
Diverse Thyroid Cancer Cell Line Culture
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THJ-11T, -16T, -21T, and -29T were all obtained from Dr. John Copland of the Mayo Clinic. U-Hth7, U-HTh74cl.7, C643, and SW1736 cell lines were obtained from Dr. Nils Erik Heldin (University of Uppsala, Sweden). Cell lines 8505C, ASH3 and KMH2 were all purchased from the Japanese Collection of Research of Bioresources Cell Bank (JCRB). Lastly, BHT-101 and CAL62 were both purchased from the DSMZ Cell Bank.
THJ-11T, -16T, -21T, and -29T cell lines were cultured in RPMI 1640 media supplemented with 10% FBS (GIBCO), 1x non-essential amino acids (Wisent), 1 mM sodium pyruvate (Wisent), penicillin (100 μg/mL) and streptomycin (100 μg/mL) (Invitrogen). U-Hth7, U-HTh74cl.7, C643, SW1736 and 8505C cell lines were cultured in EMEM media supplemented with 10% FBS (GIBCO), penicillin (100 μg/mL) and streptomycin (100 μg/mL) (Invitrogen). ASH3 and KMH2 cell lines were cultured in a 1:1 mixture of DMEM and RPMI 1640, which was supplemented with 10% heat-inactivated FBS (GIBCO), penicillin (100 μg/mL) and streptomycin (100 μg/mL) (Invitrogen). BHT-101 and CAL62 cell lines were cultured in DMEM supplemented with 10% heat-inactivated FBS (GIBCO), 1% human serum (Wisent), penicillin (100 μg/mL) and streptomycin (100 μg/mL) (Invitrogen).
THJ-11T, -16T, -21T, and -29T cell lines were cultured in RPMI 1640 media supplemented with 10% FBS (GIBCO), 1x non-essential amino acids (Wisent), 1 mM sodium pyruvate (Wisent), penicillin (100 μg/mL) and streptomycin (100 μg/mL) (Invitrogen). U-Hth7, U-HTh74cl.7, C643, SW1736 and 8505C cell lines were cultured in EMEM media supplemented with 10% FBS (GIBCO), penicillin (100 μg/mL) and streptomycin (100 μg/mL) (Invitrogen). ASH3 and KMH2 cell lines were cultured in a 1:1 mixture of DMEM and RPMI 1640, which was supplemented with 10% heat-inactivated FBS (GIBCO), penicillin (100 μg/mL) and streptomycin (100 μg/mL) (Invitrogen). BHT-101 and CAL62 cell lines were cultured in DMEM supplemented with 10% heat-inactivated FBS (GIBCO), 1% human serum (Wisent), penicillin (100 μg/mL) and streptomycin (100 μg/mL) (Invitrogen).
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