Atcc htb 38

Manufactured by American Type Culture Collection

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ATCC® HTB-38™ is a cell line derived from a human lung carcinoma. It is a commonly used in vitro model for research purposes.

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25 protocols using atcc htb 38

Cell Cultivation and Viability Assay

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HT-29 cells (ATCC^® HTB-38^™) were obtained from ATCC (Manassas, VA). Recombinant mouse hepatoma cells (H1L6.1c3) was a gift from Gregory
Kennedy (University of Alabama at Birmingham) to Jan-Peter van Pijkeren. The antioxidant response element reporter-HepG2 cell line (HepG2-ARE) designed to monitor the Nrf2 antioxidant response pathway activation was purchased from BPS Bioscience, Inc. (San Diego, USA). Cell cultures were maintained in 75-cm² tissue culture flasks in DMEM supplemented with 10 % v/v FBS, 1% v/v penicillin-streptomycin (10,000 U/ml) and incubated at 37°C with 5% CO₂ humidified atmosphere. Cells (80–90% confluence) were subcultured every 3 days, passaging at a 1:5 split ratio. Cell viability was measured using a standard MTT assay [29 (link)].

Tocmo R., Le B., Heun A., Pijkeren J.P., Parkin K, & Johnson J.J. (2020). Prenylated xanthones from mangosteen (Garcinia mangostana) activate the AhR and Nrf2 pathways and protect intestinal barrier integrity in HT-29 cells. Free radical biology & medicine, 163, 102-115.

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Cell Culture Protocol for Colon and Intestinal Cells

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Human colon cancer cells (ATCC HTB-38) and rat small intestine epithelial cells (IEC-6, ATCC CRL-1592) were purchased from the American Type Culture Collection (ATCC; Rockville, MD, USA). The cells were maintained in a humidified 5% CO₂, 95% air, 37°C environment in Roswell Park Memorial Institute (RPMI)-1640 medium. Dulbecco’s modified Eagle’s medium (DMEM) was supplemented with penicillin/streptomycin (P/S), and HT-29 and IEC-6 cell cultures were supplemented with 10% fetal bovine serum (FBS; HyClone, Inc., South Logan, UT, USA). Cells were cultured to 80% confluency in 100-mm dishes. The medium was replaced every 2 days.

KANG M.H., KIM I.H, & NAM T.J. (2014). Phloroglucinol induces apoptosis via apoptotic signaling pathways in HT-29 colon cancer cells. Oncology Reports, 32(4), 1341-1346.

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Culturing Authenticated HT-29 Cells

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Authenticated, mycoplasma-free HT-29 cells (ATCC® HTB38™), a human carcinoma cell line with epithelial morphology were obtained directly from the American Type Culture Collection (ATCC). Morphology was routinely monitored to verify cell line authenticity and cells were periodically tested for contamination by PCR. Cells were cultured in Dulbecco’s Modified Eagle Medium GlutaMAX (DMEM) (Gibco) supplemented with 10% fetal bovine serum (FBS) (Gibco) at 37 °C in 5% CO₂–95% air. Cells were split 1:12 and passaged as the culture reached confluence. To prepare for co-culture with bacteria, HT-29 cells were detached from the growth surface with 0.25% trypsin–1 mM EDTA (Gibco), resuspended in DMEM+FBS, counted, diluted to the appropriate density in DMEM+FBS, and seeded in 96-well plates to confluence (approximately 5 × 10⁵ HT-29 cells).

Wheeler K.M., Cárcamo-Oyarce G., Turner B.S., Dellos-Nolan S., Co J.Y., Lehoux S., Cummings R.D., Wozniak D.J, & Ribbeck K. (2019). Mucin glycans attenuate the virulence of Pseudomonas aeruginosa in infection. Nature microbiology, 4(12), 2146-2154.

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Establishing Colorectal Cancer Spheroids

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Human colorectal cancer cell lines HCT-116 and HT-29 were selected because of their reported sensitivity towards Artesunate [61 (link),62 (link)] and their spheroid formation ability [63 (link),64 (link),65 (link)]. Cell lines were purchased from ATCC (HCT-116, ATCC CCL-247; HT-29, ATCC HTB-38) and cultured according to the manufacturer’s instructions. Cell line-derived cancer spheroids were prepared as described previously [63 (link)]. Briefly, monolayer cultures with a confluency of at least 90% were detached with 0.05% trypsin/0.53 mM EDTA (Pan Biotech, Aidenbach, Germany). Cell number and viability were determined using the trypan blue exclusion assay. Cell suspensions with a viability of at least 90% were used for spheroid formation. 5 × 10⁴ vital cells were seeded in each well of a low adhesive 96-well microtiter plate and incubated for 48 h under standard culture conditions (37 °C, 5% CO₂). A single cancer spheroid was obtained in each well. Cancer cell lines were used for a maximum of 10 passages. Mycoplasma assays were performed quarterly.

Niederreiter M., Klein J., Arndt K., Werner J, & Mayer B. (2023). Anti-Cancer Effects of Artesunate in Human 3D Tumor Models of Different Complexity. International Journal of Molecular Sciences, 24(9), 7844.

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HT-29 Colorectal Cancer Cell Culture

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The human colorectal carcinoma cell line HT-29 (ATCC^® HTB-38™), purchased from ATCC (American Type Cell Collection), was cultured in specific medium: McCoy’s 5a modified medium (ATCC^® 30-2007™) completed with 10% fetal bovine serum (FBS, Gibco) and 1% of antibiotic mixture (penicillin and streptomycin, Sigma Aldrich, Merck KGaA Darmstadt, Germany). Standard conditions were implemented for cell culturing: a temperature of 37 °C and 5% CO₂ in a humidified incubator.
Aspartame powder, penicillin and streptomycin mixture, phosphate-buffered saline (PBS), trypsin EDTA, Trypan blue, and Alamar blue (resazurin) were acquired from Sigma Aldrich, Merck KGaA. The cell culture specific media—McCoy’s 5a Medium Modified (ATCC^® 30-2007™) and fetal bovine serum (FBS, Gibco)—were purchased from ATCC and Thermo Fisher Scientific, Inc., Waltham, MA, USA, respectively. All the reagents were of analytical standard purity and were applied according to the manufacturers’ recommendations.

Maghiari A.L., Coricovac D., Pinzaru I.A., Macașoi I.G., Marcovici I., Simu S., Navolan D, & Dehelean C. (2020). High Concentrations of Aspartame Induce Pro-Angiogenic Effects in Ovo and Cytotoxic Effects in HT-29 Human Colorectal Carcinoma Cells. Nutrients, 12(12), 3600.

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Caco-2 and HT-29 Cell Line Culture

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Caco-2 (ATCC^® HTB-37™) and HT-29 (ATCC^® HTB-38™) cell lines were obtained from ATCC, LGC Standards, Middlesex, UK and authenticated by short tandem repeat (STR) profiling (Eurofins Genomics, Ebersberg, Germany). Cells were cultured in high-glucose (4500 mg/L) Dulbecco’s modified Eagle’s media (DMEM) Gibco^® Invitrogen™ Life Technologies Ltd., Paisley, UK, containing 10% foetal bovine serum (FBS), 4 mM L-glutamine and a mixture of 50 IU/mL penicillin and 50 μg/mL streptomycin. In addition, Caco-2 cells were supplemented with 0.1 mM MEM NEAA (non-essential amino acids). Cell lines were incubated at 37 °C, with 5% carbon dioxide (CO₂) and constant humidity.

Brice D.P., Murray G.I., Wilson H.M., Porter R.J., Berry S., Durum S.K, & McLean M.H. (2022). Interleukin-27 Regulates the Function of the Gastrointestinal Epithelial Barrier in a Human Tissue-Derived Organoid Model. Biology, 11(3), 427.

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Culturing Authenticated HT-29 Cells

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Culturing Gastric, Colorectal Cancer Cells

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Gastric adenocarcinoma (AGS; ATCC CRL-1739) and carcinoma (NCI-N87; ATCC CRL-5822) cells, colorectal adenocarcinoma (HT-29; ATCC HTB-38) and carcinoma (HCT-116; ATCC CCL-247) cells were purchased from ATCC (Manassas, VA, USA). AGS cells were maintained in F-12K medium (ATCC), NCI-N87 were maintained in RPMI-1640 medium, HT-29 and HCT 116 cells were maintained in McCoy’s 5A modified medium. All the culture media were complexed with 10% fetal bovine serum (ATCC). All the cells tested were placed at 37 °C in a humidified atmosphere of 5% CO₂ to retain the proliferating condition.

Russo C., Barone D., Lavorgna M., Piscitelli C., Macaluso M., Pacifico S., Piccolella S., Giordano A, & Isidori M. (2022). Cytotoxic evaluation and chemical investigation of tomatoes from plants (Solanum lycopersicum L.) grown in uncontaminated and experimentally contaminated soils. Scientific Reports, 12, 13024.

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Cytotoxicity Assay of Viruses on Cancer Cell Lines

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HT-29 (ATCC^® HTB-38^™, American Type Culture Collection, Manassas, VA, USA) colorectal carcinoma cell line [61 (link)] was selected for the in vitro cytotoxicity study of ECHO-7 virus. The HT-29 colorectal carcinoma cell line was grown in Dulbecco’s modified eagle medium (DMEM) supplemented with 10% fetal calf serum (FCS), 1% penicillin-streptomycin (PS) and 2 mM glutamine cultured at 37 °C and 5% CO₂ incubator.
LNCaP metastatic prostate cancer cell line was provided by Professor Magnus Essand [62 (link)] and selected for the in vitro cytotoxicity study of adenovirus Ad[I/PPT-E1A]. The LNCaP metastatic prostate cancer cell line was grown in Roswell Park Memorial Institute (RPMI) 1640 supplemented with 10% fetal calf serum (FCS) and 1% penicillin-streptomycin (PS) cultured at 37 °C and 5% CO₂ incubator. Cells were routinely tested for mycoplasma and authenticated using PCR.

Deng S., Iscaro A., Zambito G., Mijiti Y., Minicucci M., Essand M., Lowik C., Muthana M., Censi R., Mezzanotte L, & Di Martino P. (2021). Development of a New Hyaluronic Acid Based Redox-Responsive Nanohydrogel for the Encapsulation of Oncolytic Viruses for Cancer Immunotherapy. Nanomaterials, 11(1), 144.

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Bacterial Adhesion to HT-29 Cells

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Adhesion was measured as previously [18] with some modification [19, 20] . Briefly, freshly late-log phase grown NCFM (20 h, OD 0.5 for tannic acid; 24 h, OD Imaging Multi-Mode Reader, BioTek) using 485 nm and 538 nm as excitation and emission wavelength, respectively. Adhesion was expressed as percentage of fluorescence recovered from the lysed bacteria that were bound versus the fluorescence of the total bacterial suspension added to the wells. Three independent experiments were conducted, each in quadruplicate, and data were subjected to one-way ANOVA using OriginPro ver. 9. Human colonic HT-29 cell line (American Type Culture Collection, ATCC® HTB38™) was cultured and maintained according to the supplier's instructions. HT-29 cells in complete growth medium containing McCoy's 5a medium (ATCC) supplemented with 10% heatinactivated fetal bovine serum (Sigma-Aldrich) and 1% penicillin-streptomycin (Lonza) were seeded in 24-well plates and cultivated until a confluent state, followed by removing the medium and washing with PBS to remove remaining antibiotics. Labeled NCFM (500 µL, OD 600 0.5) was added to the wells (2 h) and adhesion analyzed as above.

Celebioglu H.U., Delsoglio M., Brix S., Pessione E, & Svensson B. (2018). Plant Polyphenols Stimulate Adhesion to Intestinal Mucosa and Induce Proteome Changes in the Probiotic Lactobacillus acidophilus NCFM. Molecular nutrition & food research, 62(4).

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