The human oral squamous cell carcinoma lines SAS, HSC-3, HSC-4, and OSC-19 were obtained from JCRB Cell Bank. All of these cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM) (Thermo Fisher Scientific, Waltham, MA) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 1% penicillin-streptomycin. They were cultured in an atmosphere of 5% CO2 at 37 °C.
Hsc 4
Manufactured by Japanese Collection of Research Bioresources Cell Bank
Sourced in Japan
The HSC-4 is a piece of laboratory equipment used for the isolation and cultivation of hematopoietic stem cells. It provides a controlled environment for the growth and maintenance of these specialized cells, which are critical for various research and clinical applications.
Automatically generated - may contain errors
Lab products found in correlation
Hsc 3, by Japanese Collection of Research Bioresources Cell Bank (25 mentions)
Hsc 2, by Japanese Collection of Research Bioresources Cell Bank (17 mentions)
Ca9 22, by Japanese Collection of Research Bioresources Cell Bank (9 mentions)
Ho 1 u 1, by Japanese Collection of Research Bioresources Cell Bank (8 mentions)
Scc 4, by Japanese Collection of Research Bioresources Cell Bank (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (5 mentions)
Ho 1 n 1, by Japanese Collection of Research Bioresources Cell Bank (4 mentions)
Osc 19, by Japanese Collection of Research Bioresources Cell Bank (3 mentions)
Hsc 4, by Nacalai Tesque (3 mentions)
Oral keratinocyte medium, by ScienCell (3 mentions)
Dulbecco s modified eagle s medium, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Nichirei Biosciences (2 mentions)
Scc 9, by Merck Group (2 mentions)
Hsc 4, by Merck Group (2 mentions)
Scc 25, by Merck Group (2 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (2 mentions)
Scc25, by American Type Culture Collection (2 mentions)
Scc 4, by Nacalai Tesque (2 mentions)
Kosc2, by Thermo Fisher Scientific (2 mentions)
39 protocols using hsc 4
1
Culturing Human Oral Cancer Cell Lines
2
Cultivation of Oral Squamous Cell Carcinoma
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The use of a human cell line was approved by the Human Research Ethics Committee, Faculty of Dentistry, Chulalongkorn University (approval no. 017/2017). Human oral squamous cell carcinoma cells (HSC-4) were purchased from the Japanese Collection of Research Bioresources Cell Bank, National Institutes of Biomedical Innovation, Health and Nutrition, Japan. The cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS), 100 unit/mL penicillin, 100 μg/mL streptomycin, 250 ng/mL amphotericin B, and 2 mM L-glutamine. The cells were kept at 37 °C in a humidified 5% carbon dioxide atmosphere. Culture medium and supplements were purchased from Gibco, CA, USA.
3
HNSCC Cell Line Culture Protocol
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Six HNSCC cell lines SAS, HSC4, UMSCC22B, SCC90, HMS001 and SCC47 were cultured as previously reported. Human HNSCC cell lines: SAS and HSC4 were purchased from the JCRB Cell Bank (Osaka, Japan), UMSCC1 and UMSCC22B were obtained from University of Michigan (Courtesy Dr. Carey), 93VU147T was obtained from Dr. Hans Joenje (VU Medical Center Van der Boechorststraat, The Netherlands), HMS001 was a generous gift from Dr. William A. Michaud (Mass. General Hospital Cancer Center, Boston, MA), SCC90 was a generous gift from Dr. Camille Ragin (University of Pittsburg Cancer Institute, Pittsburgh, PA), and UM-SCC47 was purchased from EMD Millipore (Temecula, CA). SAS, HSC4, UMSCC22B, SCC90, and 147T were maintained in DMEM containing 10% fetal calf serum (FCS), penicillin (100U/ml), and streptomycin (0.1mg/ml). UMSCC1 was maintained in DMEM supplemented with 10% FBS, hydrocortisone (1μg/ml), penicillin (100U/ml), and streptomycin (0.1mg/ml). HMS001 was cultured in DMEM/F12 containing 10% FBS, penicillin (100U/ml), and streptomycin (0.1mg/ml). UM-SCC47 was maintained in DMEM high Glucose, containing 10% FCS, non-essential Amino Acids, penicillin (100U/ml), and streptomycin (0.1mg/ml). All the cells were maintained in a 5% CO2 atmosphere at 37 °C.
4
Head and neck cancer cell lines
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The HSC-2 (JCRB0622), HSC-3 (JCRB0623), and HSC-4 (JCRB0624) human head and neck squamous cell carcinoma (HNSCC) cell lines were obtained from the Japanese Collection of Research Bioresources (JCRB) cell bank (Osaka, Japan). Human gingival fibroblasts (HGF, CRL-1740) and the PC3 (CRL-1435) and LNCaP (CRL-1740) human prostate cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The cells used in this study were maintained in Dulbecco’s modified Eagle’s medium (Sigma, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS; Invitrogen, Carlsbad, CA, USA; complete DMEM) at 37 °C in a humidified atmosphere containing 5% CO2. The establishment of RANKL-expressing HSC-2 HNSCC cells was described previously18 (link). Among them, the RANKL-expressing R1 and R2 cells and the control C1 cells were used in this study.
5
OSCC Cell Line Characterization and Treatments
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The human OSCC cell lines HSC2, HSC3, HSC4, and KON were obtained from the Japanese Collection of Research Bioresources (JCRB) Cell Bank, Osaka, Japan, and SCC25 cells were purchased from the American Type Culture Collection (ATCC), Manassas, VA, USA. All cell lines were authenticated by JCRB and ATCC using short tandem repeat (STR) analysis. Total RNA from the normal tongue was purchased from Biochain (Newark, CA, USA) and used as a control. Cells were maintained in Dulbecco's modified Eagle's medium (DMEM; Wako Pure Chemical, Osaka, Japan) supplemented with 10% fetal bovine serum (Nichirei Biosciences, Tokyo, Japan) in 5% CO2 in air at 37°C. Anti-MIA antibody (R&D Systems) was used for neutralizing MIA in cultured medium at 2 μL/mL concentration. Further, 20 mM of recombinant human MIA (rhMIA) (Abnova, Taipei, Taiwan) treatment was performed. Moreover, cells were treated with 10 nM paclitaxel (Wako Pure Chemical), cisplatin (Wako Pure Chemical), and 5-FU (Wako Pure Chemical).
6
Cell line culture conditions
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The human HNSCC cell lines HSC-2, HSC-3, and HSC-4 were obtained from the Japanese Collection of Research Bioresources cell bank (Osaka, Japan). SCC-9 and SCC-25 cells were obtained from the American Type Culture Collection (Manassas, VA, USA). The prostate cancer cell line DU145 was obtained from the NCI Division of Cancer Treatment and Diagnosis (Bethesda, MD, USA). HSC-2, HSC-3, and HSC-4 cells were cultured in Dulbecco’s modified Eagle medium (DMEM; #D5796, Sigma, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS; #FBS-12A, Capricorn Scientific, Ebsdorfergrund, Germany) at 37°C in a humidified atmosphere of 5% CO2. SCC-9 and SCC-25 cells were cultured in DMEM/Nutrient Mixture F12 medium (containing L-glutamine and sodium bicarbonate without HEPES; the medium was liquid, sterile-filtered, and suitable for cell culture; DMEM/F12; #D8062, Sigma) supplemented with 10% FBS and 400 ng/mL hydrocortisone (#088-02483, Fujifilm Wako Pure Chemical Corporation, Osaka, Japan) at 37°C in a humidified atmosphere of 5% CO2. DU145 cells were cultured in RPMI medium containing 10% FBS at 37°C in a humidified atmosphere of 5% CO2.
7
OSCC Cell Line Culturing Protocol
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8
Salivary Gland Organoid Culture Protocol
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Methods of salivary gland organoid culture have been previously described (Yoshimoto et al., 2020 (link)). This study was performed with the permission of the ethics committee in Fukuoka Dental College (number 406). Salivary glands were obtained from the patients with mucous cysts or head and neck tumors who underwent an operation in Fukuoka Dental College hospital. Human squamous cell carcinoma cell line, HSC-4, was purchased from JCRB cell bank (Osaka, Japan). Spheroid culture was previously described (Yoshimoto et al., 2022 (link), 2023 (link)). Briefly, 1×104 cells in 200 μl Dulbecco's Modified Eagle's Medium (DMEM, Sigma-Aldrich, St. Loius, MO, USA) with 10% FBS were seeded in each well of 96-well Nunclon Sphera plate (Thermo Fisher Scientific, Waltham, MA, USA). 10% neutral buffered formalin or 4% paraformaldehyde -fixed and paraffin-embedded organoids tissue blocks were cut into 4-μm-thick sections and mounted on adhesive glass slides (CRE-01, Matsunami Glass, Osaka, Japan) for HE and immunohistochemical staining. Frozen sections were prepared by embedding samples in Tissue-Tek optimal-cutting-temperature (OCT) compound (Sakura Finetek Japan, Tokyo, Japan) and cryosectioning. The images of HE and immunohistochemical staining were captured using an AXIO Vert.A1 microscope (Carl Zeiss, Oberkochen, Germany).
9
Cultivation of Human HNSCC Cell Lines
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We used three cell lines established from human HNSCC (tongue SCCs): SCC4, SAS, and HSC4 from JCRB Cell Bank (Osaka, Japan). The cells were grown in a mixture of Dulbecco’s modified Eagle’s medium and Ham’s F-12 (SCC4), or RPMI1640 (SAS and HSC4), supplemented with 10 % fetal bovine serum (FBS), 1 % Antibiotic-Antimycotic mixture stock solution (100X) (Nakarai Tesque, Kyoto, Japan) in a humidified incubator (37 °C, 5 % CO2).
10
Cellular Characterization of Oral Cancer
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SAS, OSC-19, SCC-4, HSC-2, HSC-3, HSC-4, and KOSC-2 cl3-43 (KOSC2) human oral cancer cells and human embryonic kidney 293 (HEK 293) cells were purchased from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan). The Jurkat E6.1 (Jurkat) human T-cell lymphoma cell line was purchased from KAC Co., Ltd. (Kyoto, Japan). SAS, KOSC2, and Jurkat cells were grown in RPMI supplemented with 10% FBS (Thermo Fisher Scientific, Waltham, MA, USA). Anti-GART rabbit monoclonal antibody (H8132) for western blot analysis and anti–β-actin mouse monoclonal antibody (ab6276) were purchased from Abcam (Cambridge, UK). Anti-GART mouse monoclonal antibody (4D6-1D5) (Abnova, Taipei, Taiwan) for immunofluorescence cytochemistry, anti-BTK mouse monoclonal antibody (ab54319), and anti-Bmx goat polyclonal (sc-8874) (Santa Cruz Biotechnology, Dallas, TX, USA) antibodies were purchased from the suppliers indicated. ITK inhibitor (Cmpd-5) was obtained from Carna Biosciences, Inc. (Hyogo, Japan) (Figure S1 ).
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