A new workflow combining magnetic cell separation and impedance-based cell dispensing for gentle, simple and reliable cloning of specific CD8+ T cells.

Details

Serval ID
serval:BIB_97892CDB341E
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
A new workflow combining magnetic cell separation and impedance-based cell dispensing for gentle, simple and reliable cloning of specific CD8+ T cells.
Journal
SLAS technology
Author(s)
Ben Khelil M., Aeberli L., Perchaud M., Genolet R., Abdeljaoued S., Borg C., Binda D., Harari A., Jandus C., Muller G., Loyon R.
ISSN
2472-6311 (Electronic)
ISSN-L
2472-6303
Publication state
Published
Issued date
04/2022
Peer-reviewed
Oui
Volume
27
Number
2
Pages
130-134
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
Reverse immunology has open the door to innovative cancer immunotherapy strategies such as immunogenic antigen-based vaccination and transgenic T cell receptor (TCR)-based adoptive cell transfer. This approach enables the identification of immunogenic tumor specific antigen derived peptides. One of the major challenges is the rapid selection of antigen-specific CD8+ T cell clones. Thus, IFNγ-producing CD8+ T cells magnetic sorting combined with limiting dilution cloning approach represents the most common method of specific T cell cloning. However, during plate setup several wells will not contain T cells whereas others will contain mixed population of T cells. In this case, a re-cloning step is required which make limiting dilution based cloning a laborious, inefficient, expensive and a time-consuming method. To address these obstacles, here we present a novel 2-step workflow combining simple, affordable and gentle magnetic cell separation followed by single cell isolation using a device called DispenCell-S1. We aimed to compare this new workflow with the traditional limiting dilution method using in vitro generated antigen-specific CD8+ T cells. Herein, we reported the reliability of DispenCell-S1 method and its efficiency in T cell clones isolation.
Keywords
CD8-Positive T-Lymphocytes, Cell Separation, Cloning, Molecular, Electric Impedance, Reproducibility of Results, Workflow, Impedance, Reverse immunology, Single cell, Specific T cell, T cell Clones
Pubmed
Web of science
Open Access
Yes
Create date
31/01/2022 11:15
Last modification date
15/07/2022 5:35
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