Presentation Title

Assessing cytokine supplementation to improve lymphocyte viability after cryopreservation.

Presenter Information

Bella CircoFollow

Advisor Information

Dr. Paul W Denton

Location

MBSC Ballroom - Poster #507 - U

Presentation Type

Poster

Start Date

4-3-2022 12:30 PM

End Date

4-3-2022 1:45 AM

Abstract

A primary goal of the Denton Immunobiology Lab is improving the killing efficacy of natural killer (NK) cells. We measure increases in the killing efficacy of NK cells enriched from human PBMCs (peripheral blood mononuclear cells). Our lab’s PBMC-based work has proven to be most effective in the killing assays when the blood cells are collected from fresh blood and used without cryopreservation. This brings challenges, associated with the constraints of not being able to freeze cells for later use, that we need to alleviate. To do this, I hypothesize that recently thawed NK cells need exogenous stimulation with cytokines to stay alive in post-thawing incubations. The first two cytokines I plan to test in this context are interleukin-15 and interleukin-2. My aim in this project is to establish lab standard concentrations of exogenous cytokine supplementation to be used when incubating recently thawed PBMCs. The goal is to identify cytokine levels that sustain the viability of the NK cells but do not, by themselves, lead to NK cell activation. My presentation will outline my experimental plans for testing the stated hypothesis. In addition, I will present work that I have performed over the past months in the lab where I was tasked with optimizing fluorescent staining conditions for our target cells in our flow cytometric NK cell killing assays.

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COinS
 
Mar 4th, 12:30 PM Mar 4th, 1:45 AM

Assessing cytokine supplementation to improve lymphocyte viability after cryopreservation.

MBSC Ballroom - Poster #507 - U

A primary goal of the Denton Immunobiology Lab is improving the killing efficacy of natural killer (NK) cells. We measure increases in the killing efficacy of NK cells enriched from human PBMCs (peripheral blood mononuclear cells). Our lab’s PBMC-based work has proven to be most effective in the killing assays when the blood cells are collected from fresh blood and used without cryopreservation. This brings challenges, associated with the constraints of not being able to freeze cells for later use, that we need to alleviate. To do this, I hypothesize that recently thawed NK cells need exogenous stimulation with cytokines to stay alive in post-thawing incubations. The first two cytokines I plan to test in this context are interleukin-15 and interleukin-2. My aim in this project is to establish lab standard concentrations of exogenous cytokine supplementation to be used when incubating recently thawed PBMCs. The goal is to identify cytokine levels that sustain the viability of the NK cells but do not, by themselves, lead to NK cell activation. My presentation will outline my experimental plans for testing the stated hypothesis. In addition, I will present work that I have performed over the past months in the lab where I was tasked with optimizing fluorescent staining conditions for our target cells in our flow cytometric NK cell killing assays.