8305C was purchased from Sigma Aldrich (Darmstadt, Germany) and cultured in RPMI with 20% fetal bovine serum (FBS), 1% L-glutamine and 1% antibiotics (100 IU penicillin and 100 μg/mL streptomycin) and 1% non-essential amino acids (NEAA). SW1736 was purchased from CLS Cell line service GmbH (Eppelheim, Germany) and cultured in RPMI 1640 medium with the above additives. MDA-T32 was purchased from ATCC (American Type Culture Collection, Manassas, VA, USA) and cultured in RPMI 1640 (Biowest, Nuaillé, France). The BHT-101 cell line was purchased from Deutsche Sammlung von Microorganismen und Zellkulturen (DSMZ GmbH, Braunschweig, Germany) and cultured in Dulbecco’s MEM (DMEM) with 20% FBS and 1% antibiotics (100 IU penicillin and 100 μg/mL streptomycin). All additives were acquired from Biochrom Kg, Berlin, Germany. All cells were incubated at 37 °C in an atmosphere containing 5% CO2.
Mda t32
Manufactured by American Type Culture Collection
Sourced in United States
The MDA-T32 is a piece of laboratory equipment used for the cultivation and maintenance of cell cultures. It provides a controlled environment with adjustable temperature, humidity, and atmospheric composition to support the growth and proliferation of various cell types.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (7 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Tpc 1, by American Type Culture Collection (3 mentions)
Rpmi 1640, by Biowest (1 mentions)
Tpc 1, by Merck Group (1 mentions)
Pcdna3.1 vector, by GenePharma (1 mentions)
Mda t32, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Mda t41, by American Type Culture Collection (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Ht ori3, by American Type Culture Collection (1 mentions)
Roswell park memorial institute (rpmi), by Avantor (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Mda t68, by American Type Culture Collection (1 mentions)
Rpmi 1640, by American Type Culture Collection (1 mentions)
Mda t85, by American Type Culture Collection (1 mentions)
Mycoscope kit, by Genlantis (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Nthy ori 3 1, by European Collection of Authenticated Cell Cultures (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
11 protocols using mda t32
1
Cell Culture Protocols for Cancer Research
2
Culturing Thyroid Cell Lines
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3
Transfection of PTC Cell Lines with DLG1-AS1 and miR-199a-3p
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IHH-4 and MDA-T32 (ATCC, USA) two human PTC cell lines were included. The cell culture medium for both cell lines was prepared by mixing DMEM and FBS with a ratio of 9:1. Cell culture conditions were 95%, 37 °C, and 5% CO2. DLG1-AS1 expression vector was constructed using pcDNA3.1 vector (GenePharma, Shanghai, China). Scramble microRNA mimics were designed as negative control microRNA (NC) and miR-199a-3p were both from RIBOBIO77 (Guangzhou, China). At confluence of 70–80, IHH-4 and MDA-T32 cells were harvested and counted, followed by the transfection of 40 nM miRNA (NC miRNA as NC group) or 10 nM vector (empty vector as NC group) into 3× 106 cells through transient transfections mediated by lipofectamine 2000 (Invitrogen, USA). Control (C) cells for all groups were untransfected cells. All subsequent experiments were performed using cells harvested at 24 h post-transfection.
4
Characterization of ATC and PTC Cell Lines
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The study included five TC-derived cell lines (MDA-T32, MDA-T41, U-hth-74, U-hth-104, and SW1736). ATC-derived cell lines U-hth-74, U-hth-104 and SW1736 were obtained from Dr. N-E Heldin and cytogenetically characterized [30 (link)]. Short tandem repeats (STR) genotyping for ATC-derived cell lines U-hth-74, U-hth-104 and SW1736 was recently performed and matched to previously published genotypes [9 (link)], while the PTC-derived cell lines MDA-T32 and MDA-T41 were purchased from ATCC in 2018. Cells were cultured in RPMI-1640 medium (Thermo Fisher Scientific, Waltham, MA) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA), 100 U/mL penicillin, 100 μg/mL streptomycin, and 4 mM L-glutamine.
5
ITGA3 Expression in Papillary Thyroid Cancer
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To verify the expression of ITGA3 in papillary thyroid cancer, normal cell line Nthy-ori 3-1 and papillary thyroid cancer cell line MDA-T32 were obtained from ATCC. These cell lines were cultured in Dulbecco's modified Eagle's medium (Gibco, USA) with 10 % fetal bovine serum (Gibco, USA) under the atmosphere at 37 °C with 5 % CO2.
6
HHLA2 Regulation in PTC Cell Lines
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Four types of human PTC cell lines (TPC-1, IHH-4, CGTH-W3, and MDA-T32) and a normal thyroid cell line (HTORI3) were purchased from ATCC and cultured in the RPMI 1640 medium (Life Technologies, USA) with 10% FBS (Life Technologies, USA). HHLA2 was silenced by the RNAi approach with the small hairpin RNA (shRNA) and was overexpressed by the transfection of lentivirus containing HHLA2 sequences. The transfection efficiency was evaluated by detecting the expression of HHLA2.
7
Thyroid Cancer Cell Line Cultivation
8
MDA-T Cell Line Cultivation
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MDA-T32, -T41, -T68, and -T85 were all purchased from American Type Culture Collection (ATCC) in December 2018. Cells were grown in RPMI-1640 (ATCC 30-2001™) supplemented with 10% FBS, 1% NEAA, and 1% L Glutamine (except MDA-T85 – as directed by ATCC). Cell lines were utilized at low passage and were routinely assayed for mycoplasma contamination with MycoScope Kit (Genlantis, San Diego, CA).
9
Culturing Thyroid Cancer Cell Lines
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Three human thyroid cancer cell lines (TPC1, MDA-T32, and 8505C) and one normal thyroid cell line (Nthy-ori-3-1) were used in this study. TPC1, MDA-T32 (American Type Culture Collection, cat#30-2001, Manassas, VA, USA), and normal cell line Nthy-ori-3-1 (European Collection of Authenticated Cell Cultures (ECACC), Porton Down, UK) were routinely cultured in RPMI1640 containing 10% fetal bovine serum (FBS), 100 units/mL penicillin, and 2 mM l -glutamine (Gibco, Carlsbad, CA, USA) at 37 °C in a humidified chamber containing 5% CO2. The 8505C cells (ECACC, Porton Down, Porton, UK) were routinely cultured in MEM containing 10% fetal bovine serum (FBS), 100 units/mL penicillin, and 1% sodium pyruvate.
10
Characterization of Thyroid Cell Lines
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Nthy-ori 3-1 is an immortalized thyroid follicular epithelial cell line derived from normal adult thyroid tissue, and Nthy-ori 3-1 cell clones expressing either wild-type BRAF (Nthy/WT) or mutant BRAF (Nthy/V600E) were established (Kim et al. 2017) (link). BCPAP and MDA-T32, the human PTC cell lines harboring both BRAF V600E and TERT promoter mutations, were kindly provided by Dr Minho Shong (Chungnam National University, Daejon, Korea) and purchased from the American Type Culture Collection, respectively. KTC-2, FRO, SW1736 and 8505C are the human anaplastic thyroid cancer cell lines harboring BRAF V600E and TERT promoter mutations. The former two cell lines were kind gifts from Dr June-Key Chung (Seoul National University College of Medicine, Seoul, Korea), and SW1736 and 8505C cells were kindly provided by Dr Yoon Woo Koh (Yonsei University College of Medicine, Seoul, Korea) and Dr Seong Jin Lee (Hallym University College of Medicine, Chuncheon, Korea), respectively. KTC-2 cells were cultured in the RPMI 1640 medium supplemented with 5% of fetal bovine serum and all other cells were cultured in the RPMI 1640 medium supplemented with 10% of fetal bovine serum, and grown at 37°C in a humidified atmosphere containing 5% of CO 2 .
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