Tib 152

Manufactured by American Type Culture Collection

Sourced in United States

TIB-152 is a cell line derived from human embryonic kidney cells. It is commonly used in various cell culture and biological research applications.

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Jurkat (TIB-152), (14 citations) ATCC® TIB-152™), (7 citations) Jurkat E6–1 (TIB-152), (3 citations) Jurkat cells (Clone E6-1 #TIB-152 (2 citations) Jurkat cells (TIB‐152) (2 citations)

76 protocols using tib 152

Diverse Cell Line Cultivation Protocol

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Human breast cancer MCF-7 cells, colorectal cancer HT-29 cells, HUVECs (passage 1; CRL-1730™) and human lung fibroblast MRC-5 cells were obtained from ATCC. EMEM (MilliporeSigma) was used to culture MCF-7, HT-29 and MRC-5 cells, whereas HUVECs and Jurkat T cells (TIB-152™; ATCC) were cultured in EBM2 (Lonza Group Ltd.) and RPMI-1640 (MilliporeSigma), respectively. All media were supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and 2 mM glutamine (MilliporeSigma). Cells were cultured at 37°C and 8% CO₂ was supplied to cells under static conditions. Two tumor cell lines (MCF-7 and HT-29) were used to show the versality of the AXTEX-4D platform.

Baru A., Mazumder S., Kundu P., Sharma S., Purakayastha B.P., Khan S., Gupta R, & Arora N.M. (2022). Establishment of a three-dimensional triculture model on the novel AXTEX-4D™ platform. Oncology Reports, 49(1), 2.

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Quantifying Anti-toxin Antibody Activity

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The anti-tHla antibody levels and functional activity (neutralizing activity, NA) of the antibodies in serum were assessed respectively by ELISA and Toxin Neutralisation Assay (TNA) as described by Oscherwitz and Cease^{33 (link)}. In brief, the ability of antibody to block recombinant alpha toxin (AT) cytotoxicity in vitro was assessed using the Jurkat T cell line (TIB-152, ATCC, Manassas, VA). Mouse anti-Staphylococcal alpha hemolysin mAb (8B7) (IBT Bioservices # 0210-001) was used as standard positive to obtain minimum and maximum levels for neutralisation of AT (H9395, Sigma Biologicals).

Diemen P.M., Leneghan D.B., Brian I.J., Miura K., Long C.A., Milicic A., Biswas S., Rollier C.S, & Wyllie D.H. (2017). The S. aureus 4-oxalocrotonate tautomerase SAR1376 enhances immune responses when fused to several antigens. Scientific Reports, 7, 1745.

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Dielectric Spectroscopy of Primary Chondrocytes and Jurkat Cells

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Dielectric spectroscopy experiments were performed on primary costal chondrocytes and a T-cell leukemia-derived Jurkat E6-1 clone cell line (ATCC^® TIB-152^™, Manassas, VA, USA). The chondrocyte cells were cultured in Chondrocyte Growth Medium (CGM; PromoCell, Heidelberg, GER), and Jurkat cells in RPMI 1640 medium (Atlanta Biologicals, Norcross, GA). RPMI and CGM supplemented with 10% fetal bovine serum from Atlanta Biologicals and PromoCell, respectively. Both mediums were also supplemented with 2 mM L-glutamine (Gibco/Invitrogen, Carlsbad, CA), 50 IU/ml penicillin (Gibco/Invitrogen), and 50 mg/ml streptomycin (Gibco/Invitrogen) at 37°C with 5% CO2 in air. All the cells were suspended in an isotonic buffer consisting of 229 mM sucrose, 16 mM glucose, 1 μM CaCl₂, and 5 mM Na₂HPO₄ in double distilled water (pH 7.4) for the experiments, after a washing step with the isotonic buffer. The measurements were performed directly after the suspension of cells in LCB. The most common medium used for DEP manipulation in the field is an isotonic sucrose/dextrose solution supplemented with minimal amount of salts for buffering (please see supplementary information S1.4), which could justify our selection of LCB.

Sabuncu A.C., Asmar A.J., Stacey M.W, & Beskok A. (2015). DIFFERENTIAL DIELECTRIC RESPONSES OF CHONDROCYTE AND JURKAT CELLS IN ELECTROMANIPULATION BUFFERS. Electrophoresis, 36(13), 1499-1506.

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Cell Culture Protocols for U87, Jurkat, and H9

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U87 cells (HTB-14; ATCC) were cultured in Dulbecco’s Modified Eagle Medium (DMEM; Gibco) supplemented with 1% GlutaMax (Gibco), 10 mM HEPES (Gibco) and 10% fetal bovine serum (FBS; Peak Serum) and passaged prior to formation of neurospheres. Jurkat T cells (TIB-152; ATCC) and H9 cells (HTB-176; ATCC) were cultured in RPMI 1640 (Gibco) supplemented with 2 mM L-glutamine (Cellgro), 25 mM HEPES (Irvine Scientific), and 10% heat-inactivated fetal bovine serum (FBS; Hyclone).

López C.L., Brempelis K.J., Matthaei J.F., Montgomery K.S., Srinivasan S., Roy D., Huang F., Kreuser S.A., Gardell J.L., Blumenthal I., Chiefari J., Jensen M.C., Crane C.A, & Stayton P.S. (2021). Arming Immune Cell Therapeutics with Polymeric Prodrugs. Advanced healthcare materials, 11(9), e2101944.

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Culturing Cell Lines and Mouse Models

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Cell lines Jurkat (ATCC® TIB-152™), Raw-Lucia ISG (http://www.invivogen.com/raw-lucia-isg), and B16F10 (ATCC® CRL-6475™), were cultured following vendor instructions (37 °C, 5% CO₂). Female C57BL/6J (JAX Stock No. 000664) mice 6–8 weeks of age were maintained in the animal facility at the Massachusetts Institute of Technology (MIT). All animal studies and procedures were carried out following federal, state, and local guidelines under an IACUC-approved animal protocol (the MIT Committee on Animal Care (CAC) Protocol Number: 0717-076-20).

Li Y., Teague B., Zhang Y., Su Z., Porter E., Dobosh B., Wagner T., Irvine D.J, & Weiss R. (2019). In vitro evolution of enhanced RNA replicons for immunotherapy. Scientific Reports, 9, 6932.

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Jurkat and Primary T Cell Labeling and Transduction

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The Jurkat T cell line was obtained commercially (#TIB-152, ATCC, Manassas, VA) for initial nanoemulsion cell labeling characterizations. Jurkat cells were grown in RPMI-1640 media (Gibco, Waltham, MA) plus 10% fetal bovine serum (FBS), 10 mM HEPES buffer (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), 1 mM sodium pyruvate and 1.5 mg/mL sodium bicarbonate.
Primary human T cells were obtained from blood samples sourced from the San Diego Blood Bank and enriched for T cells by Ficoll (Histopaque 1077, Sigma Aldrich) gradient density centrifugation and magnetic assisted cell sorting (Dynabeads, Thermo Fisher). T-cells were then activated with human T-activator CD3/CD28 Dynabeads and allowed to expand for two days in RPMI-1640 supplemented with 10% FBS and 100 units/mL of recombinant human interleukin 2 (IL-2, Peprotech, Rocky Hill, NJ). For transduction, we employed a vector specific to epidermal growth factor receptor variant III (EGFR-vIII) as described by Johnson et al. (30 ). A detailed CAR virus production and human T cell transduction is available elsewhere (31 (link)). CAR receptor expression was confirmed by flow cytometry. We used T cell populations with >70% CAR+ expression.

Hingorani D.V., Chapelin F., Stares E., Adams S.R., Okada H, & Ahrens E.T. (2019). Cell penetrating peptide functionalized perfluorocarbon nanoemulsions for targeted cell labeling and enhanced fluorine-19 MRI detection. Magnetic resonance in medicine, 83(3), 974-987.

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Galectin-9 Apoptosis Inhibition Assay

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Example 12

The ability of Galectin-9 G9.2-17 and G9.1-8m9 antibodies to prevent Galectin-9-induced apoptosis of Jurkat cells was assessed. Jurkat cells (TIB-152, ATCC, Manassas, Va.) were grown in modified RPMI (2 mM L-glutamine, 10 mM HEPES, 1 mM sodium pyruvate, 4.5 g/L glucose, 1.5 g/L sodium bicarbonate) with 10% FBS, 100 mU/mL penicillin and 100 μg/mL streptomycin at 37° C. with 5% CO₂. Cells (2×10⁵cells/well) were incubated in the wells of a 96-well culture plate with or without the addition of 280 nM Galectin-9 (2045-GA, RnD Systems, Minneapolis, Minn.), 1 μM G9.2-17 IgG, and/or 1 μM G9.1-8m9 IgG. Cells were incubated at 37° C., 5% CO₂for 5 hours and then resuspended in annexinV-binding buffer. Cells were then stained with AnnexinV-AlexaFluor488 (Invitrogen, Carlsbad, Calif.) at 4° C. for 30 minutes in the dark. Prior to flow cytometry, propidium iodide (PI)_ was added (1 μg/mL). Cells were run on Guava easyCyte flow cytometer (MilliporeSigma, Burlington, Mass.) and analyzed using FlowJo (Treestar, Ashland, Oreg.). Cell population was gated via forward and side scatter, then analyzed on AnnexinV for all apoptotic cells and PI for late apoptotic cell populations. Results are shown in FIG. 23.

US11414492B2. Anti-galectin-9 antibodies and uses thereof (2022-08-16). New York University [US], PureTech Health, LLC [US]. Inventors: Shohei Koide [US], George Miller [US], Akiko Koide [US], Linxiao Chen [US], Eric Elenko [US], Aleksandra Filipovic [GB], Joseph Bolen [US].

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Culturing Human Osteoblasts and T-lymphocytes

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Human bone tissue-derived osteoblasts (CRL-11372) [American Type Culture Collection (ATCC), Manassas, VA, USA] were maintained in the logarithmic phase of growth in a 1:1 mixture of Ham’s F12 medium/Dulbecco’s modified Eagle’s medium (Gibco-BRL, Grand Island, NY, USA), with 2.5 mM l-glutamine (without phenol red), 0.3 mg/ml G418, and supplemented with heat-inactivated 10% fetal bovine serum (Gibco-BRL) at 34°C in a 5% CO₂–95% air-humidified incubator. Human Jurkat T-lymphocyte leukemia cell lines (TIB-152; ATCC) were maintained in the logarithmic phase of growth in RPMI-1640 medium (Gibco-BRL) supplemented with heat-inactivated 10% fetal bovine serum (Gibco-BRL) at 37°C in a 5% CO₂–95% air-humidified incubator.

Ma W., Jin W., He X., Sun Y., Yin H., Wang Z, & Shi S. (2022). Mycobacterium tuberculosis Induced Osteoblast Dysregulation Involved in Bone Destruction in Spinal Tuberculosis. Frontiers in Cellular and Infection Microbiology, 12, 780272.

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Evaluating Cytotoxicity of Au/Pt/ZnO Nanoparticles

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In this work, the influence of Au/Pt/ZnO nanoparticles on the vitality of human cells was evaluated in vitro, with the use of established human Acute T Cell Leukemia cell line, Jurkat (ATCC® TIB-152™), as well as mononuclear cells isolated from peripheral blood (PBMC) of voluntary donors. PBMC contains both peripheral blood lymphocytes (PBL) and monocytes. Cells were cultured in suspension in RPMI 1640 medium with 2 mM L-glutamine with the addition of 10% fetal bovine serum (FBS) and 1% Gibco® Antibiotic-Antimycotic solution (10,000 units/mL of penicillin, 10,000 μg/mL of streptomycin, and 25 μg/mL of Amphotericin B) on 24-well plastic plates (TC-PLATE 24 well, Greiner). Cell cultures were carried out in an incubator at 37 °C in a 5% CO₂ atmosphere with increased humidity. The Au/Pt/ZnO nanoparticles have been added in three various concentrations, 1 μM, 10 μM, and 100 μM. As controls served the samples without the Au/Pt/ZnO nanoparticles addition. All tested samples, both Jurkat cells, and PBMC were cultured for 24, 48 and 72 h.

Dobrucka R., Romaniuk-Drapała A, & Kaczmarek M. (2020). Biologically synthesized of Au/Pt/ZnO nanoparticles using Arctium lappa extract and cytotoxic activity against leukemia. Biomedical Microdevices, 22(4), 72.

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Induction of Primary Cilia in RPE-1 Cells

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Telomerase-immortalized human pigment epithelial cells RPE-1 (CRL-4000; ATCC) were grown in DMEM/F12 with 10% FBS, and ECV304 cells (CRL-1998; ATCC) and Jurkat T cells (TIB-152; ATCC) in DMEM with 10% FBS. Cells were incubated at 37°C in an atmosphere of 5% CO₂/95% air. Primary cilium formation in RPE-1 cells was induced by starving the cells in DMEM/F12 with 0.25% FBS for 24 h.

Fernández-Barrera J., Bernabé-Rubio M., Casares-Arias J., Rangel L., Fernández-Martín L., Correas I, & Alonso M.A. (2018). The actin-MRTF-SRF transcriptional circuit controls tubulin acetylation via α-TAT1 gene expression. The Journal of Cell Biology, 217(3), 929-944.

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