Flotation-Based T Cell Isolation Using Buoyancy-Activated Cell Sorting

Published: July 31, 2023

Abstract

Source: Snow, T., et al. Flotation-Based T Cell Isolation, Activation, and Expansion from Human Peripheral Blood Mononuclear Cell Samples Using Microbubbles. J. Vis. Exp. (2022)

This video demonstrates T cell isolation from peripheral blood mononuclear cells (PBMCs) using the functionalized buoyancy-activated cell sorting (BACS) technology. The isolation technique involves the positive selection of CD3+ T cells by labeling the cells with biotinylated anti-CD3 antibodies and separating the antibody-bound cells using streptavidin-bound microbubbles.

Protocol

1. Isolation of T cells with microbubbles using positive selection NOTE: This protocol details a small-scale CD3+ positive selection approach using SAMBs. Incubate 3 x 108 commercially obtained PBMCs in 2.5 mL of separation buffer with biotinylated anti-CD3 (OKT3) antibody at a concentration of 25 ng of antibody per 1 million cells (25 ng/M). Gently mix by pipetting up and down, and incubate at room temperature for 10 min.</li…

開示

The authors have nothing to disclose.

Materials

Biotin anti-human CD3 (OKT3) Antibody Biolegend 317320
Leukopak BioIVT HUMANLMX100-0001129
Normal Human PBMCs BioIVT HUMANHLPB-0002562
Streptavidin Microbubble Kit (includes Akadeum's separation buffer) Akadeum 11110-000

タグ

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記事を引用
Flotation-Based T Cell Isolation Using Buoyancy-Activated Cell Sorting. J. Vis. Exp. (Pending Publication), e21528, doi: (2023).

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