Htb 177

Manufactured by American Type Culture Collection

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HTB-177 is a cell line obtained from a human breast cancer sample. It is a commonly used in vitro model for research on breast cancer biology and the development of cancer therapies.

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ATCC-HTB-177 (3 citations) H460 (HTB-177) (3 citations)

14 protocols using htb 177

Imaging of human cancer cell lines

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Four human cancer cell lines and one normal, were selected as tissues frequently subjected to imaging in radiotherapy (Table 1). The cell lines, obtained from American Type Culture Collection (ATCC) and CellBank Australia, were cultured in a humidified incubator with 5% CO₂ at 37 °C, according to the manufacturers’ instructions, and allowed to reach 80% confluence. Cell cultures were maintained at passage numbers lower than 15. No antibiotics or antifungal agents were used to minimize stress on cells [18] (link).Table 1

Human cell lines chosen for this study, the tissue of origin and the type of cancer/disease.

Cell Line	Tissue of origin	Disease
NCI-H460 (ATCC® HTB-177™)	Lung: pleural effusion	Carcinoma; large cell lung cancer
DU 145 (ATCC® HTB81™)	Prostate; derived from metastatic site: brain	Carcinoma
CAL 27 (ATCC® CRL2095™)	Tongue	Squamous cell carcinoma
Hs 683 (ATCC® HTB138™)	Brain	Glioma
PNT1A (CellBank Australia 95012614)	Prostate, human post pubertal, immortalized with SV40	Normal

Kench P.L., Rogers L., Esteves A., Gorjiara T., Mackonis E.C., Morrell S., McKenzie D.R, & Suchowerska N. (2020). Imaging prior to radiotherapy impacts in-vitro survival. Physics and Imaging in Radiation Oncology, 16, 138-143.

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Cell Lines for Cancer Research

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The human lung cancer cell lines used in the experiments were A-549 (human lung epithelial cells, ATCC ^® CCL-185™) and NCI-H460 (human lung epithelial cells, ATCC ^® HTB-177™). For the human glioblastoma cell line, U-87MG (Korea Cell Line Bank ^®, Seoul, Republic of Korea) was used. The media were from Hyclone (Cytiva, Pittsburgh, PA, USA): RPMI 1640 MEDIUM (1X) (cat. no. SH30027.01) for the lung cancer cell line and DMEM/HIGHGLUCOSE (cat. no. SH30243.01) for the glioblastoma medium. Both of these media were supplemented with 10% FBS (cat. no. SH30919.03) and 1% penicillin–streptomycin solution (cat. no. SV30010) from the same company. Cells were cultured at 37 °C with 5% CO₂ in a humidified incubator.

Kahm Y.J., Jung U, & Kim R.K. (2023). Regulation of Cancer Stem Cells and Epithelial-Mesenchymal Transition by CTNNAL1 in Lung Cancer and Glioblastoma. Biomedicines, 11(5), 1462.

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Cell Culture and Transfection of Lung Adenocarcinoma

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Lung adenocarcinoma cells A549 (CCL-185, ATCC, Manassas, VA, USA) and H460 (HTB-177, ATCC, Manassas, VA, USA) were cultured in Dulbecco’s modified eagle medium (DMEM; CAT#01–055, Biological industries, Beit HaEmek, Israel). Meanwhile, human embryonic lung fibroblasts HFL-1 (CC-Y1584, EK-Bioscience, Shanghai, China) were cultured in Ham’s F-12 (CAT#01–095, Biological industries). All media were then supplemented with 10% fetal bovine serum (FBS; GIBCO™, CAT# 10270106, Life Technologies, San Jose, CA, USA), 100 units/mL penicillin, 100 µg/mL streptomycin, and 2 mM L-glutamine, which were all purchased from Biological industries (Kibbutz Beit Haemek, Israel). Non-essential amino acids (NEAA; CAT# X0557, 1: 100, Biowest, Logan, Utah, USA) were employed for HFL-1 culture. Afterwards, the cells were cultured in a humidified incubator (Thermo Fisher Scientific Inc., Waltham, MA, USA) at 37 °C with 5% CO₂.
A549 cells were transfected with the HA-BIRC5 expression vector (pcDNA3-HA-BIRC5) and Myc-DR5 expression vector (DR5 Myc-tag) or corresponding empty vector (pcDNA) following the instructions of the Lipofectamine 2000 transfection reagent (CAT# 11668019, Invitrogen, Carlsbad, CA, USA). Cells were transfected with 1 μg of pcDNA3-HA-BIRC5 and DR5 Myc-tag or the corresponding empty vector pcDNA. After a 24-h period of transfection, the cells were treated with NPs for subsequent analyses.

Chen S., Han F., Huang D., Meng J., Chu J., Wang M, & Wang P. (2021). Fe3O4 magnetic nanoparticle-enhanced radiotherapy for lung adenocarcinoma via delivery of siBIRC5 and AS-ODN. Journal of Translational Medicine, 19, 337.

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Cytotoxicity Evaluation of Compounds

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Examined 4a–4e compounds were dissolved in DMSO (1 × 10⁻² mol dm⁻³). The experiments were carried out on 4 human cell lines. The cell lines purchased from the American Type Culture Collection (ATCC, Poland) were used: H460 (lung carcinoma, large cell lung cancer (ATCC^®HTB-177™)), MCF-7 (breast adenocarcinoma, ATCC^® HTB-22™), HCT116 (colorectal carcinoma ATCC^® CCL-247™) and HEK 293 (embryonic kidney cells); ATCC^® CRL-1573™).
Cells were cultured as monolayers and maintained in Dulbecco’s modified Eagle medium (DMEM), supplemented with 10% fetal bovine serum (FBS), 2 mM of L-glutamine, 100 U/mL of penicillin and 100 μg/mL of streptomycin in a humidified atmosphere with 5% CO₂ at 37 °C.

Mikulin I., Ljubić I., Piantanida I., Vasilev A., Mondeshki M., Kandinska M., Uzelac L., Martin-Kleiner I., Kralj M, & Tumir L.M. (2021). Polycationic Monomeric and Homodimeric Asymmetric Monomethine Cyanine Dyes with Hydroxypropyl Functionality—Strong Affinity Nucleic Acids Binders. Biomolecules, 11(8), 1075.

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NSCLC and Normal Cell Cultures

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The NSCLC A549cells (ATCC® CCL-185™) were maintained in Dulbecco´s Modified Eagle Medium/ Nutrient Mixture F-12 (DMEM/F12) supplemented with 4 mM of L-glutamine and 10% of fetal serum bovine (FBS) at 37°C in an atmosphere that contained 5% CO₂. NCI-H460 (ATCC® HTB-177™, a NSCLC) and MRC5 (ATCC® CCL171™, a normal cell line) cells were grown in Roswell Park Memorial Institute (RPMI) 1640 medium or Dulbecco’s Modified Eagle Medium (DMEM), respectively, and those cells were maintained at the same temperature and atmosphere in the presence of 10% of FBS. For experiments, cells were seeded in specific plates at the density of 3.2 x 10³ cells/ cm², and the heptapeptide ang-(1–7) (Bachem Americas Inc, USA) was added or not to the cultures at a final concentration of 10⁻⁷ M [20 (link),21 (link)]. The media was renewed every 24 h and the cells were used in the assays when the cultures reached ~90% confluency.

da Silva B.D., Lima K.F., Gonçalves L.R., da Silveira M.B, & Moraes K.C. (2016). MicroRNA Profiling of the Effect of the Heptapeptide Angiotensin-(1-7) in A549 Lung Tumor Cells Reveals a Role for miRNA149-3p in Cellular Migration Processes. PLoS ONE, 11(9), e0162094.

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3D In Vitro Lung Cancer Microtissues

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The present study was conducted using two different lung carcinoma cell lines, as follows: (i) A549 cells (CCL-185 from ATCC, LGC Standards GmbH)—lung carcinoma with an epithelial-like morphology; (ii) NCI-H460 (HTB-177 from ATCC, LGC Standards GmbH) cell line—large lung cancer cells. Both cell lines were stored under standard conditions (vapor phase of liquid nitrogen) before culturing.
The EpiAirway 3D in vitro microtissues (Air-100, Lot 33251, Kit C) were acquired from MatTek Life Science Company (Bratislava, Slovak Republic) and were handled according to the EpiAirway^TM AIR-100 Use Protocol.
More details regarding the culture conditions are presented in the Supplementary Materials.

Moacă E.A., Watz C., Faur A.C., Lazăr D., Socoliuc V., Păcurariu C., Ianoș R., Rus C.I., Minda D., Barbu-Tudoran L, & Dehelean C.A. (2022). Biologic Impact of Green Synthetized Magnetic Iron Oxide Nanoparticles on Two Different Lung Tumorigenic Monolayers and a 3D Normal Bronchial Model—EpiAirwayTM Microtissue. Pharmaceutics, 15(1), 2.

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Non-Small Cell Lung Cancer Cell Lines

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Non-small cell lung cancer cells used in the experiments including NCI-H460 [H460] (ATCC^® HTB-177™, RRID: CVCL_0459), NCI-H292 [H292] (ATCC^® CRL-1848™, RRID: CVCL_0455), NCI-H23 [H23] (ATCC^® CRL-5800™, RRID: CVCL_1547), and A549 (ATCC^® CCL-185™, RRID: CVCL_0023) were obtained from the American Type Culture Collection (Manassas, VA, USA). H460, H292, and H23 cells were grown in Roswell Park Memorial Institute (RPMI) 1640 medium, and A549 cells was grown in Dulbecco’s Modified Eagle Medium (DMEM). All cell culture mediums were supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, and 100 units/mL of each of penicillin and streptomycin. Cells were placed in a humidified atmosphere of 5% carbon dioxide (CO₂) at 37 °C.

Petsri K., Yokoya M., Tungsukruthai S., Rungrotmongkol T., Nutho B., Vinayanuwattikun C., Saito N., Takehiro M., Sato R, & Chanvorachote P. (2020). Structure–Activity Relationships and Molecular Docking Analysis of Mcl-1 Targeting Renieramycin T Analogues in Patient-derived Lung Cancer Cells. Cancers, 12(4), 875.

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Cytotoxicity of Broccoli Seedling Extract

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The in vitro cytotoxicity of broccoli seedling extract was determined on MEF (mouse embryonal fibroblasts), HaCaT (normal human keratinocytes), HepG2 (liver cells—hepatocellular carcinoma), HCT116 (colon cells—colorectal carcinoma), and H460 (lung carcinoma) cell lines using dimethylthiazol diphenyltetrazolium bromide (MTT) and neutral red uptake assay. The sources of the cell lines are as follows: MEF (established in the laboratory of Dr. Pinterić from mice strain RRID:IMSR_JAX:002448), HaCat (RRID:CVCL_0038; gift of Dr. Marijeta Kralj, Ruđer Bošković Institute, Zagreb, Croatia), HepG2 (HB-8065, ATCC, Manassas, VA, USA), HCT116 (CCL-247, ATCC, Manassas, VA, USA), and H460 (HTB-177, ATCC, Manassas, VA, USA).

Gmižić D., Pinterić M., Lazarus M, & Šola I. (2023). High Growing Temperature Changes Nutritional Value of Broccoli (Brassica oleracea L. convar. botrytis (L.) Alef. var. cymosa Duch.) Seedlings. Foods, 12(3), 582.

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Cell Culture Conditions for Cancer Cell Lines

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SUM159, HBL100, MCF7, MDA-MB-468, MDA-MB-435, MDA-MB-231, SKBR3, BT549, and 293T cells were obtained from the American Type Culture Collection (ATCC; Manassas, VA). All of the above cells grown in DMEM medium (Life Technologies) supplemented with 10% fetal bovine serum (FBS; Hyclone Logan, UT, USA) and 1% penicillin/streptomycin (Life Technologies). A549, CRL-1848^TM, CRL-5803, CRL-5866, CRL-5833, 827, PC-9, 95D, HTB-182, and HTB-177 cells were obtained from the ATCC and maintained in RPMI-1640 (Hyclone) medium. The normal breast epithelial cell line MCF10A was purchased from ATCC and cultured in DMEM/F12 medium (Life Technologies) supplemented with 5% horse serum (Life Technologies), 10 μg/mL insulin (Sigma), 100 ng/mL cholera toxin (Sigma), 20 ng/mL epidermal growth factor (EGF, R&D), 0.5 μg/mL hydrocortisone (Sigma), and 1% penicillin/streptomycin. The normal lung epithelial cell line BEAS-2B was obtained from ATCC and maintained in LHC-8 (Gibco, USA). All cell lines were grown by incubation at 37°C in a humidified 5% CO₂ atmosphere.

Song L., Chang R., Dai C., Wu Y., Guo J., Qi M., Zhou W, & Zhan L. (2016). SORBS1 suppresses tumor metastasis and improves the sensitivity of cancer to chemotherapy drug. Oncotarget, 8(6), 9108-9122.

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Cultivation of Lung Cancer and Normal Cells

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Lung cancer cell (LCC) lines H1299, H460, SPC-A1, A549, and HEK293T (CRL-5803, HTB-177, CRL-5803, CCL-185, and CRL-11268, ATCC, United States) and normal lung cells 16HBE (PCS-300-010, ATCC, United States) were purchased from ATCC. All the cells were cultured in the dulbecco’s modified eagle medium (DMEM,10566024, Gibco, United States) containing penicillin-streptomycin double antibody and 10% fetal bovine serum (FBS; 10437028, Gibco, United States) in a constant temperature 5%CO2incubatorat 37°C.

Wang R.T., Zhang Y., Yao S.Y, & Tan X.G. (2020). LINC00501 Inhibits the Growth and Metastasis of Lung Cancer by Mediating miR-129-5p/HMGB1. OncoTargets and therapy, 13, 7137-7149.

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