Hoechst 33342 stains were purchased from Life Technologies (Cat no. H3570). Human Chordoma cell lines U-CH1 (Cat. no. CRL-3217) and U-CH2 (Cat. no. CRL-3218) were purchased from ATCC (Manassas, VA, USA). Cells were grown in Iscove’s Modified Dulbecco’s Medium (Cat. no. 30-2005, ATCC): RPMI-1640 Medium (Cat. no. 30-2001, ATCC) (4:1), 10% FBS (Cat. no. 30-2020, ATCC) supplemented with 2 mM L-glutamine (Cat. no. 30-2214, ATCC). HEK-293 (Cat. no. CRL-1573) and HUVEC (Cat. no. PCS-100-013) cells were purchased from ATCC and were grown in Eagle’s Minimum Essential Medium (Cat. no. 30-2003, ATCC) supplemented with 10% fetal bovine serum, and Vascular Cell Basal Medium (Cat. no. PCS-100-030, ATCC) supplemented with Endothelial Cell Growth Kit-BBE (Cat. no. PCS-100-040, ATCC), respectively. PSMB5 (Cat. no. 11903), PSMB8 (Cat. no. 13635), and β-Actin (Cat. no. 3700) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Doxorubicin was purchased from Sigma (Cat. no. D1515). Compound libraries were provided by the Institute of Chemistry and Cell Biology (ICCB) Longwood, Harvard Medical School, Boston, MA, USA.
U ch2
Manufactured by American Type Culture Collection
Sourced in United States
The U-CH2 is a laboratory equipment product designed for cell culture applications. It serves as a compact and efficient incubator for maintaining optimal environmental conditions for cell growth and proliferation. The core function of the U-CH2 is to provide a controlled temperature, humidity, and gas atmosphere to support the cultivation of various cell types.
Automatically generated - may contain errors
Lab products found in correlation
U ch1, by American Type Culture Collection (10 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Mug chor1, by American Type Culture Collection (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640 medium, by American Type Culture Collection (2 mentions)
Two step luminescence assay, by Lonza (2 mentions)
Iscove s modified dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Doxorubicin, by Merck Group (1 mentions)
Vascular cell basal medium, by American Type Culture Collection (1 mentions)
Endothelial cell growth kit bbe, by American Type Culture Collection (1 mentions)
Psmb5, by Cell Signaling Technology (1 mentions)
Psmb8, by Cell Signaling Technology (1 mentions)
Fetal bovine serum (fbs), by American Type Culture Collection (1 mentions)
L glutamine, by American Type Culture Collection (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Hek293, by American Type Culture Collection (1 mentions)
Panc 1, by American Type Culture Collection (1 mentions)
X vivo 10 medium, by Lonza (1 mentions)
Heat inactivated human ab serum, by Omega Scientific (1 mentions)
13 protocols using u ch2
1
Chordoma Cell Line Cultivation and Compound Screening
2
Chordoma Cell Line Validation
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UM-Chor1 (clival) and JHC7 (sacral) were kindly provided by the Chordoma Foundation on approximately April 2, 2015 with short tandem repeat (STR) validation. U-CH1 (sacral), U-CH2 (sacral), UM-Chor5 (clival), and MUG-CC1 (clival) were obtained from ATCC on approximately July 20, 2020 with STR validation. All cell lines were passaged twice per week, used at a low (<30) passage number, and serially verified to be free of Mycoplasma infection using a two-step luminescence assay (Lonza Bioscience).
3
Chordoma and Notochord Cell Protocol
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This study used discarded tumor tissues from patients with chordoma as well as notochord cells from discarded human embryonic tissues. The Institutional Review Board at Massachusetts General Hospital (Boston, MA) approved the chordoma study protocol (2009A052093) and the notochord study protocol (2007P-002239). As both protocols used discarded material, the Institutional Review Board waived the need for written informed consent. Cell lines U-CH1, U-CH2, U-CH12, HEK293, and 293T were purchased from ATCC (Manassas, VA), and CH22 cells were provided by F.J.H. Cells were cultured in a biosafety level 2 environment according to ATCC guidelines.
4
Chordoma Cell Line Validation Protocol
5
Cell Line Characterization and Genetic Manipulation
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The human lung H460, ovarian ES2, pancreatic PANC-1, and chordoma U-CH1, U-CH2, and MUG-Chor1 carcinoma cell lines were obtained from American Type Culture Collection (ATCC) and maintained in culture as recommended by the ATCC. All cell lines were newly purchased or their identity confirmed by STR analysis (PANC-1 and H460 cells). Brachyury overexpression and silencing vectors and transfection strategies were previously described [14 (link)].
6
Chordoma Cell Lines and haNK Cells Protocol
7
Chordoma Cell Line CRISPR Transfection
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Human chordoma cell lines JHC7 and UCH2 were purchased from the American Type Culture Collection (Manassas, VA, USA). JHC7 and UCH2 cells were seeded at 2 × 103 cells/plate in 96-well plates, 2 × 104 cells/plate in 24-well plates, or 2 × 105 cells/plate in 6-well plates and cultured in DMEM/F12 and IMDM/RPMI 1640 (4:1) medium supplemented with 10% fetal bovine serum (FBS), respectively. After 24 h, cells were cultured with fresh warm culture medium and treated with 50 ng of Cas9/gRNA RNP VLP for an additional 48 or 72 h depending on the experiment.
8
Chordoma Cell Line Culture and Analysis
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The human chordoma cell lines U-CH1 and U-CH2 were both obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) and were cultured in a 1:4 ratio of Iscove’s modified Dulbecco’s modified Eagle’s medium (Gibco, Grand Island, NY, USA) and RPMI-1640 medium (Gibco) supplemented with 10% fetal bovine serum (FBS; Gibco) and 1% penicillin/streptomycin (Gibco) in a humidified incubator with a 5% CO2/95% air atmosphere at 37 °C. Culture flasks were coated with rat tail type I collagen (BD Biosciences, San Diego, CA, USA) prior to use. The following antibodies were used in the experiments: anti-Vimentin, anti-N-cadherin and anti-GAPDH were obtained from Cell Signaling Technology (Beverly, MA, USA), and anti-Smad3 and anti-E-cadherin were obtained from Abcam (USA). Smad3 small interfering RNA (siRNA) was purchased from Suzhou GenePharma (Suzhou, China). Lipofectamine 3000 was purchased from Origene (Rockville, MD, USA).
9
Culturing Chordoma Cell Lines
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The human chordoma cell lines U-CH1 and U-CH2 were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). Both lines were grown in a mixture of Iscove’s Modified Dulbecco’s Medium (ATCC) and RPMI-1640 Medium (Gibco; Thermo Fisher Scientific, Waltham, MA, USA) at a ratio of 1:4. This basal medium was supplemented with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc), 100 U/mL penicillin (Gibco/Thermo Fisher Scientific, Inc), 100 mg/mL streptomycin (Gibco; Thermo Fisher Scientific, Inc), and 1% L-glutamine (Gibco; Thermo Fisher Scientific, Inc). Both cell lines were cultured at 37 °C in a humidified atmosphere containing 5% CO2.
10
Culture of Chordoma and HEK-293T Cell Lines
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Human chordoma cell lines (U-CH1 and U-CH2) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and incubated in Iscove’s modified Dulbecco’s modified Eagle’s medium (Invitrogen, Carlsbad, CA, USA) and RPMI-1640 medium (Invitrogen) at a ratio of 1:4, supplemented with 10% fetal bovine serum (FBS, Invitrogen) and 1% penicillin/streptomycin (Beyotime, Haimen, Jiangsu, China). Human embryonic kidney 293T (HEK-293T) cells were also obtained from ATCC and cultured in RPMI-1640 medium containing 10% FBS and 1% penicillin/streptomycin. The incubator was maintained at 37°C and 5% CO2.
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