Cell Regeneration

Cell Regeneration

Volume 7, Issue 1, September 2018, Pages 7-15
Cell Regeneration

A protocol for generating induced T cells by reprogramming B cells in vivo

https://doi.org/10.1016/j.cr.2018.05.001Get rights and content
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Abstract

Obtaining T cells by reprogramming is one of the major goals in regenerative medicine. Here, we describe a protocol for generating functional T cells from Hoxb5-expressing pro/pre-B cells in vivo. This protocol includes the construction of Hoxb5 recombinant plasmids, retroviral packaging, isolation and viral transduction of pro/pre-B cells, cell transplantation, and phenotypic analysis of induced T cells. The procedure is reproducible and straightforward, providing an approach for generating induced T cells for translational research.

Keywords

Hoxb5
Retrovirus
Pro/pre-B cells
T cells

Abbreviations

CAR-T
Chimeric antigen receptor T-Cell Immunotherapy
TAA-TCR-T
tumor-associated antigen-TCR-T
HSC
Hematopoietic stem cells
SP
single positive
iPSC
induced pluripotent stem cells
GFP
green fluorescent protein
PB
peripheral blood
LN
Lymph node
DN
double negative
PE
Phycoerythrin
APC
Allophycocyanin
BV
Brilliant Violet
PerCP
Peridinin Chlorophyll
DAPI
4,6-diamidino-2-phenylindole
FITC
Fluorescein isothiocyanate
7-AAD
7-Aminoactinomycin D

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Peer review under responsibility of Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences.