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Celltrace cfse proliferation kit

Manufactured by Thermo Fisher Scientific
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The CellTrace CFSE Proliferation Kit is a fluorescent cell staining solution used for tracking cell division and proliferation in vitro. The kit contains the CFSE (Carboxyfluorescein succinimidyl ester) dye, which covalently binds to intracellular proteins, allowing the labeling and monitoring of cell division through flow cytometry or microscopy.

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Lab products found in correlation

Cellevent caspase 3 7, by Thermo Fisher Scientific (2 mentions) Facscalibur, by BD (2 mentions) Cd3 cd28 dynabead, by Thermo Fisher Scientific (1 mentions) Eclipse ti e inverted microscope, by Nikon (1 mentions) A1r confocal laser scanning system, by Nikon (1 mentions) Nis elements software, by Nikon (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by AppliChem (1 mentions) Anti cd3 cd28 coated dynabeads, by Thermo Fisher Scientific (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Easysep human na ve cd4 t cell enrichment kit, by STEMCELL (1 mentions) Cell proliferation dye efluor 670, by Thermo Fisher Scientific (1 mentions) Facsaria 2 flow cytometer, by BD (1 mentions) Nivolumab, by MedChemExpress (1 mentions) Magnetic separation column, by Miltenyi Biotec (1 mentions) Macsquantify, by Miltenyi Biotec (1 mentions) Aim 5 medium, by Thermo Fisher Scientific (1 mentions) Cd8 percp cy5, by BioLegend (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Anti cd3, by Miltenyi Biotec (1 mentions) Anti cd28, by BD (1 mentions)

Close spelling variants of this product

CellTrace CFSE Cell Proliferation Kit CellTrace CFSE or Violet (CTV) Cell Proliferation Kit CellTrace™ carboxyfluorescein succinimidyl ester (CFSE) cell proliferation kit CellTrace™ CFSE Cell Proliferation Kit for flow cytometry CellTrace™ CFSE Cell Proliferation Kit Protocol CellTrace CFSE kit CellTrace Proliferation Kit CellTrace CFSE CFSE kit

24 protocols using celltrace cfse proliferation kit

1

T cell and NK cell activation protocols

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HD T cells were stained using the CellTrace CFSE Proliferation Kit (Thermo Fisher Scientific, C34554) according to the manufacturer's instructions. For stimulation, CD3/CD28 Dynabeads (Thermo Fisher Scientific, 11131D) at a bead-to-cell ratio of 1:2 and 30 U/mL interleukin-2 (IL-2; R&D Systems, 202-IL-010/CF) were added to the culture for 5 days. T cell subsets were discriminated by the expression of the chemokine receptor CCR7 in combination with the naïve cell marker CD45Ra (Supplementary Figure 2). NK cells were stimulated by addition of 500 U/mL IL-2 for 10 days. Fold change was calculated as: “Viable NK cell count day 10”/“Viable NK cell count day 0.”
Rohrbacher L., Brauchle B., Ogrinc Wagner A., von Bergwelt-Baildon M., Bücklein V.L, & Subklewe M. (2021). The PI3K∂-Selective Inhibitor Idelalisib Induces T- and NK-Cell Dysfunction Independently of B-Cell Malignancy-Associated Immunosuppression. Frontiers in Immunology, 12, 608625.
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2

Cell Proliferation and Apoptosis Assay

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Cells were plated on six-well plates at a density of 5 × 104 for HeLa or 1 × 105 for A549 cells, and counted every 24 h for three days when grown either at 21% or 1% O2. Cells were counted using a hemocytometer in triplicate. Cell division was also assessed using both CFSE assay (CellTrace CFSE Proliferation kit, Thermo Scientific). Cell death via apoptosis was measured using a caspase 3 activity kit (CellEvent Caspase 3/7, Thermo Fischer), according to manufacturer’s instructions.
Ortmann B.M., Burrows N., Lobb I.T., Arnaiz E., Wit N., Bailey P.S., Jordon L.H., Lombardi O., Peñalver A., McCaffrey J., Seear R., Mole D.R., Ratcliffe P.J., Maxwell P.H, & Nathan J.A. (2021). The HIF complex recruits the histone methyltransferase SET1B to activate specific hypoxia-inducible genes. Nature genetics, 53(7), 1022-1035.
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3

Proliferation Assay of hCB-CD34+ T Cells

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Human T cells, isolated from the spleens of (hCB)-CD34+ NSG mice, were stained using CellTrace™ CFSE Proliferation kit (ThermoFisher, Cat. No C34554), according to manufacturer’s protocol. Four days post culture, proliferation of these cells was assessed via FACS.
Tanaskovic O., Verga Falzacappa M.V, & Pelicci P.G. (2019). Human cord blood (hCB)-CD34+ humanized mice fail to reject human acute myeloid leukemia cells. PLoS ONE, 14(9), e0217345.
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4

Imaging CAR T Cells Interacting with GUVs

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On the day of the experiment, BFP-sorted cells were stained with a green CellTrace™CFSE proliferation kit (Thermo Fisher Scientific) following the manufacturer's instructions and rested in standard culture conditions, for at least 2 h, before starting the experiment. Cells were pre-washed with and kept in HBSS supplemented with 1% (v/v) FCS, 1 mM l-glutamine, 1% (v/v) non-essential amino acids (NEAA), 10 mM N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (HEPES), 0.55% (w/v) d-glucose while imaging. GUVs were added to the CAR T cell samples and imaged at 37 °C using an incubator stage (Okolab) mounted onto a confocal laser scanning microscope (Nikon Eclipse Ti-E inverted microscope equipped with Nikon A1R confocal laser scanning system, 60 × oil immersion objective, NA = 1.49, with three lasers: 488 nm, 561 nm, and 640 nm). Image acquisition and processing were performed using the NIS-Elements software (version 4.5, Nikon). The assembly of the videos (Online Resource 1–2) was done with Fiji Image J 2.1.0 software.
Meléndez A.V., Velasco Cárdenas R.M., Lagies S., Strietz J., Siukstaite L., Thomas O.S., Tomisch J., Weber W., Kammerer B., Römer W, & Minguet S. (2022). Novel lectin-based chimeric antigen receptors target Gb3-positive tumour cells. Cellular and Molecular Life Sciences, 79(10), 513.
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5

Cytotoxic Assay of NK Cell Therapies

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SS- and PROD-produced UTD-NK cells as well as SS-/PROD-produced CAR-NK cells were co-incubated for four hours with OCI-AML2 cells at an effector-to-target (E:T) ratio of 1:5. OCI-AML2 cells were previously labeled using Cell Trace CFSE proliferation kit (Thermo Fisher Scientific, Waltham, MA, USA). Subsequently, cells were stained with 4′,6-diamidino-2-phenylindole (DAPI) (AppliChem, Darmstadt, Germany) and the viability of target cells was analyzed using a BD FACSCelestaTM device (BD Biosciences).
Albinger N., Müller S., Kostyra J., Kuska J., Mertlitz S., Penack O., Zhang C., Möker N, & Ullrich E. (2024). Manufacturing of primary CAR-NK cells in an automated system for the treatment of acute myeloid leukemia. Bone Marrow Transplantation, 59(4), 489-495.
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6

T-cell Proliferation Assay on MSCs

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To assess proliferation, sorted T cells, labeled with CellTrace CFSE proliferation kit (Thermo Fisher Scientific, MA, USA), were stimulated with anti-CD3/CD28-coated Dynabeads (Thermo Fisher) at 1:1 ratio and cultured on a monolayer of MSCs, as described above, for 72 h. T cells, harvested and surface stained for CD4 surface marker, were acquired using an LSR II flow cytometer (BD Biosciences). The proliferation index was calculated as total number of divisions divided by the number of cells that underwent division using FlowJo software (BD Biosciences).
Cho W.J., Mittal S.K, & Chauhan S.K. (2023). Mesenchymal Stromal Cells Suppress T-Cell-Mediated Delayed-Type Hypersensitivity via ALCAM-CD6 Interaction. Stem Cells Translational Medicine, 12(4), 221-233.
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7

Cell Proliferation and Apoptosis Assay

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Cells were plated on six-well plates at a density of 5 × 104 for HeLa or 1 × 105 for A549 cells, and counted every 24 h for three days when grown either at 21% or 1% O2. Cells were counted using a hemocytometer in triplicate. Cell division was also assessed using both CFSE assay (CellTrace CFSE Proliferation kit, Thermo Scientific). Cell death via apoptosis was measured using a caspase 3 activity kit (CellEvent Caspase 3/7, Thermo Fischer), according to manufacturer’s instructions.
Ortmann B.M., Burrows N., Lobb I.T., Arnaiz E., Wit N., Bailey P.S., Jordon L.H., Lombardi O., Peñalver A., McCaffrey J., Seear R., Mole D.R., Ratcliffe P.J., Maxwell P.H, & Nathan J.A. (2021). The HIF complex recruits the histone methyltransferase SET1B to activate specific hypoxia-inducible genes. Nature genetics, 53(7), 1022-1035.
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8

Evaluation of T Cell Responses to DC Stimuli

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T cell proliferation and IFNγ secretion assays were performed following co-culture of CFSE labeled allogeneic naïve CD4+ T cells with sCD40L-exposed DCs +/- stimulated with Poly-ICLC as described above. Naïve CD4+ T cells were purified by magnetic cell sorting (EasySep™ Human Naïve CD4+ T cell Enrichment Kit, Stem Cell Technologies, Vancouver, Ca) from uninfected donor PBMCs and stained with 10 μM carboxyfluorescein succinimidyl ester dye (CellTrace™ CFSE Proliferation Kit, Life Technologies, NY, USA) for 10 minutes prior to incubation with DCs at a ratio of 1:20 (DC/Tcell). After 5-6 days, CFSE dilution was analyzed by FACS and culture supernatants were assayed for IFNγ using the Human Th1/Th2/Th17 Cytokine Cytometric Bead Array (CBA) (BD Pharmingen). Regulatory T cell phenotype was analyzed after co-culturing allogeneic naïve CD4+ T cells with sCD40L-exposed DCs at a ratio of 1:10 for 7 days. Cells were stained with CD4 (BD biosciences), FoxP3 (eBioscience), and CD127 (Biolegend) according to the manufacturer's instructions.
MILLER E.A., GOPAL R., VALDES V., BERGER J.S., BHARDWAJ N, & O'BRIEN M.P. (2015). Soluble CD40 ligand contributes to dendritic cell mediated T-cell dysfunction in HIV-1 infection. AIDS (London, England), 29(11), 1287-1296.
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9

Isolation and Transfer of TRP2-specific CD8+ T Cells

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Spleens from unimmunized TRP2 TCR transgenic mice were collected and dissociated as described above to obtain a single-cell suspension. The cells were treated with erythrocyte lysis buffer (water, 0.15 M NH 4 Cl, 1 mM KHCO 3 , 0.1 mM EDTA; 1 mL per spleen) at room temperature (RT) for 3 min and washed with ice-cold PBS. If the cell pellet was red after centrifugation, the lysis step was repeated. If tracing of the transferred cells was required, cells were labeled with CFSE (CellTrace CFSE Proliferation Kit, Life Technologies, Thermo Fisher Scientific) according to the manufacturer's instructions. A sample of the splenocytes was stained with the H-2K b -TRP2 180-188 tetramer and antibodies against surface markers as described above to determine the frequency of TRP2-specific CD8 + T cells. Total cell number was counted using a hemocytometer and the splenocytes were administered intravenously at 5 Â 10 5 TRP2-specific CD8 + T cells per recipient.
Purde M.T., Cupovic J., Palmowski Y.A., Makky A., Schmidt S., Rochwarger A., Hartmann F., Stemeseder F., Lercher A., Abdou M.T., Bomze D., Besse L., Berner F., Tüting T., Hölzel M., Bergthaler A., Kochanek S., Ludewig B., Lauterbach H., Orlinger K.K., Bald T., Schietinger A., Schürch C., Ring S.S, & Flatz L. (2024). A replicating LCMV-based vaccine for the treatment of solid tumors. Molecular therapy : the journal of the American Society of Gene Therapy, 32(2).
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10

HUVEC Proliferation Assay with Cigarette and E-Cigarette Extracts

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The proliferation of HUVECs was analysed using the CellTrace CFSE Proliferation Kit according to the manufacturer`s instructions (LifeTechnologies). Briefly, following staining of cells with the dye carboxyfluorescein diacetate succinimidyl ester (CFSE) and rinsing of non-bound dye, 1 x 105 stained cells were seeded into each well of a 6 well plate. Following an overnight incubation and replacement of culture medium with fresh medium, cells were treated with the cigarette smoke- or e-cigarette vapour-extracts as indicated. After treatment periods of 24 hours (data not shown) and 48 hours, cells were washed, trypsinized, and subjected to CFSE quantification on a FACS Calibur by determination of mean fluorescence intensities of the viable cells. Mean fluorescence intensity of controls was set at 100% and the different incubations were calculated in relation to the corresponding control.
Putzhammer R., Doppler C., Jakschitz T., Heinz K., Förste J., Danzl K., Messner B, & Bernhard D. (2016). Vapours of US and EU Market Leader Electronic Cigarette Brands and Liquids Are Cytotoxic for Human Vascular Endothelial Cells. PLoS ONE, 11(6), e0157337.
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