Vybrant carboxyfluorescein diacetate succinimidyl ester cfda se cell tracer kit

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom

The Vybrant Carboxyfluorescein Diacetate Succinimidyl Ester (CFDA SE) Cell Tracer Kit is a fluorescent cell labeling agent. It is used to track and monitor cell populations.

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2 protocols using vybrant carboxyfluorescein diacetate succinimidyl ester cfda se cell tracer kit

Isolation and CFSE-Labeling of G9 CD8+ T cells

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G9Cα-/-NOD transgenic mouse spleens were harvested and processed into a single cell suspension using a tissue homogenizer (a total of 53 mouse spleens were used in these studies). Following osmotic lysis of the erythrocytes, the splenocyte suspension was filtered through a 40μm cell strainer and centrifuged. CD8 + T cells were isolated by negative selection using the MACS CD8a + T cell isolation kit, according to the manufacturer's instructions. Using the Vybrant Carboxyfluorescein Diacetate Succinimidyl Ester (CFDA SE) Cell Tracer Kit (Thermo Fisher Scientific, UK) the isolated G9 CD8 + T cells were incubated with 2μM CFDA SE with the final cell suspension being produced in sterile saline (sodium chloride 0.9%) to a concentration of 4x10 7 /ml. CFDA SE is converted intracellularly into fluorescent CFSE and the CFSE-labelled G9 CD8 + T cells can be detected by flow cytometry.

Arikat F., Hanna S.J., Singh R.K., Vilela L., Wong F.S., Dayan C.M., Coulman S.A, & Birchall J.C. (2020). Targeting proinsulin to local immune cells using an intradermal microneedle delivery system; a potential antigen-specific immunotherapy for type 1 diabetes. Journal of controlled release : official journal of the Controlled Release Society, 322.

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In Vivo Cytotoxicity Assay for HPV16 E7

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Seven days after the last vaccination, mice (5/group) were injected intravenously under inhalation anesthesia with 10×10⁶ CFSE-labeled naïve splenocytes that were pulsed with 0.5 μg/mL HPV16 E7_(49–57) peptide or an irrelevant control peptide. E7-pulsed target cells were labeled with 10 μM CFSE (Vybrant Carboxyfluorescein diacetate succinimidyl ester (CFDA SE) Cell Tracer Kit, Thermo Fisher Scientific, Carlsbad, CA). Control peptide-pulsed cells were labeled with 0.66 μM CFSE. After extensive washing in PBS, cells were mixed in a 1:1 ratio prior to injection. Twenty hours after injection into vaccinated mice, splenocytes were isolated and analyzed by flow cytometry. Percentage of remaining CFSE^hi and CFSE^low cells were analyzed using FlowJo flow cytometry analysis software (Becton, Dickinson & Company, Ashland, OR). Five thousand CFSE⁺ events were collected. Peptide specific lysis was calculated with reference to the loss of the CFSE^hi target cell population using the following calculation: Percent Specific Lysis = [(%CFSE^hi−%CFSE^low)]/(%CFSE^hi)] × 100.

DA SILVA D.M., SKEATE J.G., CHAVEZ-JUAN E., LÜHEN K.P., WU J.M., WU C.M., KAST W.M, & HWANG K. (2019). Therapeutic efficacy of a human papillomavirus type 16 E7 bacterial exotoxin fusion protein adjuvanted with CpG or GPI-0100 in a preclinical mouse model for HPV-associated disease. Vaccine, 37(22), 2915-2924.

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