As previously described 29 (link), the human HNC cell lines HN4, HN6 and HN30 were kindly provided by the University of Maryland Dental School, USA. SCC-4, SCC-9, SCC-25 and CAL-27 cells were purchased from the American Type Culture Collection (ATCC, USA). The human HB cell line was derived from HIOEC by treatment with benzo[a]pyrene 30 (link) and the Rca-T cell line was purified from tongue squamous cell carcinoma tissues induced by adding 4-nitroquinoline-1-oxide into Sprague-Dawley rats' drinking water 31 (link). CAFs and normal fibroblasts (NFs) were isolated from tumor and adjacent normal tissues of HNC patients by primary culture and were identified by the presence of CAF-specific markers (α-SMA, Vimentin). All these cells except SCC-4, SCC-9 and SCC-25 were cultured in Dulbecco's modified Eagle's medium (DMEM; GIBCO-BRL, USA) supplemented with 10% heat-inactivated FBS (GIBCO-BRL), penicillin (100 units/mL), and streptomycin (100 μg/mL) at 37°C in a humidified 5% CO2 atmosphere, while SCC-4, SCC-9 and SCC-25 cells were maintained in DMEM/F12 medium containing 10% FBS. In addition, normal primary head and neck epithelial cells were cultured in keratinocyte serum-free medium (KSF; GIBCO-BRL, USA) with 0.2 ng/mL recombinant epidermal growth factor (rEGF; Invitrogen, USA).
Scc 4
Manufactured by Thermo Fisher Scientific
Sourced in United States
The SCC-4 is a laboratory instrument designed for concentration and evaporation of liquid samples. It features a compact design and can accommodate up to four samples simultaneously.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (22 mentions)
Cal27, by American Type Culture Collection (13 mentions)
Scc25, by Thermo Fisher Scientific (10 mentions)
Scc25, by American Type Culture Collection (8 mentions)
Dmem f12, by Thermo Fisher Scientific (7 mentions)
Scc 4, by American Type Culture Collection (6 mentions)
Cal27, by Thermo Fisher Scientific (6 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (6 mentions)
Hydrocortisone, by Merck Group (5 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (4 mentions)
Scc 9, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (2 mentions)
Bovine pituitary extract, by Thermo Fisher Scientific (2 mentions)
Hsc 3, by American Type Culture Collection (2 mentions)
Scc15, by American Type Culture Collection (2 mentions)
Schneider s drosophila medium, by Thermo Fisher Scientific (2 mentions)
Skn 3, by Japanese Collection of Research Bioresources Cell Bank (2 mentions)
Skn 3, by Thermo Fisher Scientific (2 mentions)
Hsc 3, by Thermo Fisher Scientific (2 mentions)
25 protocols using scc 4
1
Establishing Head and Neck Cancer Cell Lines
2
Human Oral Cancer Cell Line Cultivation
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Human OSCC cell lines H357, SCC-4 and SCC-9 were obtained from Sigma-Aldrich (collected from European Collection of Cell Cultures). The human pharynx squamous cell carcinoma cell line FaDU was obtained from the American Type Culture Collection. SCC-4 and SCC-9 cell lines were cultured in Dulbecco's Modified Eagle's Medium/F12 (DMEM/F12; Life Technologies) supplemented with 10% Fetal Bovine Serum (FBS), 0.4 μg/ml hydrocortisone (Sigma-Aldrich) and 0.5 mM sodium pyruvate (Life Technologies). FaDU was cultured in Eagle's Minimum Essential Medium (Life Technologies) supplemented with 10% FBS. H357 cells were cultured in DMEM/F12 medium supplemented with 10% FBS and 0.5 μg/ml sodium hydrocortisone succinate (Sigma-Aldrich). Primary Human Oral Keratinocytes (HOK) were isolated from healthy gingival tissue of normal human patients and maintained in keratinocyte serum-free Media (Life Technologies) supplemented with 2% FBS, bovine pituitary extract 60 mg/mL (Life Technologies) and epidermal growth factor (1 ng/mL) (Life Technologies) as described previously [34 (link)].
3
Cell Line Sources for Oral Cancer Research
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Dr. Cheng-Chia Yu (Taiwan, Chung Shan Medical University) generously provided the following cell lines: The Smulow–Glickman (S-G) human gingival epithelial cell line was original from F.H. Kasten, East Tennessee State University, Quillen College of Medicine, Johnson City, TN [12] (link); OC2 is a cell line derived from an oral squamous cell carcinoma specimen of a buccal-mucosa squamous carcinoma from a Chinese man. OC2 cell line was original from R.C. Chang [13] (link); SCC-4, a human tongue squamous cell carcinoma, was obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan) [14] (link); SAS, a high-grade tumorigenic human tongue squamous cell carcinoma, was obtained from the Japanese Collection of Research Bioresources (Tokyo, Japan) [15] (link); and SASVO3, a highly malignant head and neck squamous cell carcinoma (HNSCC) cell line obtained from primary SAS tumors using three sequential rounds of xenotransplantation [16] (link). SG, SAS, and SASVO3 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 2 mM glutamine. SCC-4 was cultured in DMEM supplemented with the nutrient mixture Ham's F-12 (Life Technologies, Grand Island, NY), 10% FBS, 2 mM glutamine, and 400 ng/mL hydrocortisone. OC2 was cultured in RPMI medium with 10% FBS in a humidified 5% CO2 atmosphere at 37°C.
4
Culturing Human Oral Cancer Cell Lines
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Human oral cancer cell lines SCC-4, SCC-9, SCC-15, SCC-25, and HSC-3, originally isolated from oral cancer (OSCC) of the human tongue, were obtained from the American Type Culture Collection (ATCC). The SCC-4, -9, -15, and -25 cells were grown in DMEM plus Ham’s F12 medium (DMEM/F-12), supplemented with 10% fetal bovine serum (FBS, Life Technologies, Inc.), penicillin (100 U/ml), streptomycin (100 μg/ml), and 0.4 μg/ml hydrocortisone. HSC3 human oral cancer cell line was placed in DMEM supplemented with 10% FBS, 100 units/ml penicillin, 100 μg/ml streptomycin, and 2 mM glutamine. The HPSE1-silenced (HPSE1-) and -upregulated (HPSE+) clones derived from the SCC-9 line were cultured in the same maintenance medium with the addition of 0.25 μg/ml puromycin and 5 μg/ml geneticin, respectively, for these cell expansion. Non-transformed human gingival keratinocyte cell line (HGK), obtained from mucosal keratinocytes and found to be spontaneously immortalized, was cultured in a serum-free, low-calcium medium (Gibco’s Keratinocyte-SFM; Invitrogen, United States) containing specific supplements and antibiotics, as previously described (Mäkelä et al., 1998 (link); Rodrigues et al., 2017 (link)). Cells were maintained at 37°C in a humidified atmosphere of 5% CO2.
5
Comparison of NSD1 Mutant and Wild-Type Cell Lines
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Three NSD1 wild-type patient-derived cell lines–Cal27 (ATCC, CRL-2095), FaDu (ATCC, HTB-43), and Detroit562 (ATCC, CCL-138) and three NSD1 mutant cell lines—SCC-4 (ATCC, CRL-1624), SKN-3 (JCRB Cell Bank, JCRB1039), BICR 78 (ECACC, 04072111) were used in this study (key resources table and Table S1 ). FaDu, Cal27, Detroit 562, SKN-3, and SCC-4 cells were cultured in Dulbecco’s modified Eagle medium (DMEM:F12; Invitrogen) with 10% fetal bovine serum (FBS; ThermoFisher). BICR78 (ECACC) was cultured in DMEM:F12 (Invitrogen) with 10% FBS (ThermoFisher), and 400ng/ml hydrocortisone (SigmaAldrich). Drosophila S2 cells were cultured in Schneider’s Drosophila medium (ThermoFisher) containing 10% heat-inactivated FBS. All cell lines tested negative for mycoplasma contamination.
6
Comparison of NSD1 Mutant and Wild-Type Cell Lines
7
Head and Neck Squamous Cell Carcinoma Cell Lines
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Cal27, HN6, HN4, HN30, SCC4, SCC9, SCC25, Fadu, and HSC3 human head and neck squamous cell carcinoma cells were purchased from the American Type Culture Collection (USA), and the mouse 4NQO-induced tongue cancer cell line MTCQ1 (BCRC 50620) was obtained from the Bioresource Collection and Research Center. Cal27, HN6, HN4, HN30, FaDu, HSC3, and MTCQ1 cell lines were maintained in DMEM (Invitrogen) containing 10% FBS (ScienCell, San Diego, CA, USA) and 1% penicillin/streptomycin (NCM Biotech, Suzhou, China) at 37 °C in a 5% CO2-humidified incubator. The SCC4, SCC9, and SCC25 were maintained in F12 (Invitrogen) containing 10% FBS (ScienCell), 1% penicillin/streptomycin (NCM biotech, China), 0.5% sodium pyruvate (Sigma-Aldrich) and 400 ng/mL Hydrocortisone (Sigma-Aldrich) at 37 °C in a 5% CO2-humidified incubator. Moreover, the three inhibitors used to cell lines were NE inhibitor (80 µM, Abcam), CG inhibitor (10 µM, Abcam), and MMP9 inhibitor (25 µM, Abcam).
8
Culturing human OSCC cell lines
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Human OSCC cell lines CAL27, HSC-2, and SCC-4 were obtained from ATCC (Manassas, VA, USA). Cells were grown in Dulbecco’s Modified Eagle’s Medium (Invitrogen, Waltham, MO, USA) for CAL27 and Minimum Essential Eagle’s Medium (Sigma-Aldrich, St. Louis, MO, USA) for HSC-2 and SCC-4 cells, supplemented with 10% fetal bovine serum (FBS) (Invitrogen) and 100 U/mL penicillin and 100 μg/mL streptomycin (Sigma-Aldrich, St. Louis, MO, USA) at 37 °C with 5% CO2.
9
OSCC and Adjacent Tissue Collection
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One-hundred and nine pairs of primary OSCC and adjacent noncancerous tissues were collected from the Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Wenzhou Medical University (Wenzhou, People’s Republic of China) between January 2005 and December 2008. All tissues were immediately frozen in liquid nitrogen and stored at −80°C until analysis. None of the patients had undergone chemotherapy or radiotherapy before surgery. The patients’ information is summarized in Table 1 . All of the patients received follow-up. Overall survival was defined as the time from primary surgery to death of the patient or, for living patients, the date of last follow-up. This study was approved by the Research Ethics Committee of the hospital, and all patients gave informed consent.
Human OSCC cell lines (SCC-4, SCC-9, SCC-25, and Tca8113), and a human normal oral keratinocyte cell line were obtained from the Beijing Institute for Cancer Research (Beijing, People’s Republic of China) and maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with Invitrogen® 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin. All the cells were incubated at 37°C in a humidified atmosphere with 5% CO2.
Human OSCC cell lines (SCC-4, SCC-9, SCC-25, and Tca8113), and a human normal oral keratinocyte cell line were obtained from the Beijing Institute for Cancer Research (Beijing, People’s Republic of China) and maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with Invitrogen® 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin. All the cells were incubated at 37°C in a humidified atmosphere with 5% CO2.
10
Cell Culture Protocols for Cancer Research
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