RT Journal Article
SR Electronic
T1 Label-free lymphocytes reconstitution using side scatter for optimal T cell manufacturing
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.11.09.375584
DO 10.1101/2020.11.09.375584
A1 Tongjin Wu
A1 Yen Hoon Luah
A1 Yongqiang Luo
A1 Howard John Womersley
A1 Lih Feng Cheow
YR 2020
UL http://biorxiv.org/content/early/2020/11/10/2020.11.09.375584.abstract
AB Lymphocyte biology research commonly involves purification of lymphocyte subpopulations by fluorescence-activated cell sorting (FACS) or immunomagnetic separation (IMS), both of which typically rely on antibody labeling of validated cell markers. Methods enabling label-free segregation of lymphocyte subpopulations would be invaluable with regard to less-perturbation, simplicity and cost-effectiveness. Here, we introduce TRuST, a label-free approach for T cell reconstitution using side-scatter (SSC). TRuST-sorted SSClow cells enrich for CD4+ T and naïve T cells, while SSChigh cells enrich for CD8+ T, NK and differentiated T cells. Enrichment purity can be improved by computational gate design. SSClow cells have superior expansion capacity and generate more central memory precursors with naïve-resembling cytokine responses. Moreover, we find that both T cell differentiation status and CD4/CD8 T ratio in the starting cellular material are critical attributes predicting T cell product quality and quantity. TRuST presents an effective and reliable technique for label-free lymphocytes selection and reconstitution.Competing Interest StatementThe authors have declared no competing interest.