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Related Experiment Video

Updated: Aug 14, 2025

Flotation-Based T Cell Isolation, Activation, and Expansion from Human Peripheral Blood Mononuclear Cell Samples Using Microbubbles

Flotation-Based T Cell Isolation, Activation, and Expansion from Human Peripheral Blood Mononuclear Cell Samples Using Microbubbles

Published on: December 23, 2022

Flotation-Based T Cell Isolation, Activation, and Expansion from Human Peripheral Blood Mononuclear Cell Samples

Tiffany Snow¹, Jon Roussey¹, Casey Wegner²

¹Research and Development, Akadeum Life Sciences.

Journal of Visualized Experiments : Jove

|January 9, 2023

Summary

A novel method uses functionalized microbubbles to isolate and activate T cells from peripheral blood mononuclear cells (PBMCs) ex vivo. This technique promotes T cell proliferation with reduced exhaustion, benefiting research and cell therapies.

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Last Updated: Aug 14, 2025

Flotation-Based T Cell Isolation, Activation, and Expansion from Human Peripheral Blood Mononuclear Cell Samples Using Microbubbles

Flotation-Based T Cell Isolation, Activation, and Expansion from Human Peripheral Blood Mononuclear Cell Samples Using Microbubbles

Published on: December 23, 2022

Isolation and Activation of Murine Lymphocytes

Isolation and Activation of Murine Lymphocytes

Published on: October 30, 2016

Assembly and Operation of an Acoustofluidic Device for Enhanced Delivery of Molecular Compounds to Cells

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Published on: January 21, 2021

Area of Science:

Immunology
Cell Biology
Biotechnology

Background:

Isolating T cells from peripheral blood mononuclear cells (PBMCs) is essential for various research and clinical applications.
Existing methods for T cell isolation and expansion can lead to cell exhaustion, limiting their utility.

Purpose of the Study:

To present a simple, novel protocol for the isolation, activation, and expansion of T cells from PBMCs ex vivo.
To utilize functionalized buoyancy-activated cell sorting (BACS) technology for improved T cell culturing.

Main Methods:

Positive selection of CD3+ cells from leukopak-derived PBMCs.
Co-stimulation using pre-conjugated anti-CD28-bound streptavidin microbubbles (SAMBs) for 48 hours.
Utilized functionalized microbubbles for buoyant activation, promoting proliferation with reduced cell contact time.

Main Results:

Isolated and cultured T cells exhibited sufficient activation and proliferation.
The method prevented overactivation and subsequent exhaustion, indicated by minimal PD-1 expression.
Functionalized microbubbles reduced T cell exhaustion by limiting co-stimulatory factor contact.

Conclusions:

The presented protocol offers an effective approach for ex vivo T cell isolation, activation, and expansion.
Buoyancy-activated cell sorting with functionalized microbubbles is a promising technique for generating robust T cell cultures.
This method supports the development of cell-based therapies and research requiring ex vivo T cell expansion.