Antibody-Based CAR T Cells Produced by Lentiviral Transduction.
Antibodies, Monoclonal
/ genetics
Biomarkers
Genetic Engineering
Genetic Vectors
/ genetics
Humans
Immunomagnetic Separation
/ methods
Immunophenotyping
Immunotherapy, Adoptive
/ methods
Lentivirus
/ genetics
Lymphocyte Activation
/ immunology
Receptors, Antigen, T-Cell
/ genetics
Receptors, Chimeric Antigen
/ genetics
T-Lymphocytes
/ immunology
Transduction, Genetic
CAR T cells
chimeric antigen receptor
lentiviral transduction
Journal
Current protocols in immunology
ISSN: 1934-368X
Titre abrégé: Curr Protoc Immunol
Pays: United States
ID NLM: 9101651
Informations de publication
Date de publication:
03 2020
03 2020
Historique:
entrez:
10
3
2020
pubmed:
10
3
2020
medline:
9
6
2021
Statut:
ppublish
Résumé
One promising approach to treat hematologic malignancies is the usage of patient-derived CAR T cells. There are continuous efforts to improve the function of these cells, to optimize their receptor, and to use them for the treatment of additional types of cancer and especially solid tumors. In this protocol, an easy and reliable approach for CAR T cell generation is described. T cells are first isolated from peripheral blood (here: leukoreduction system chambers) and afterwards activated for one day with anti-CD3/CD28 Dynabeads. The gene transfer is performed by lentiviral transduction and gene transfer rate can be verified by flowcytometric analysis. Six days after transduction, the stimulatory Dynabeads are removed. T cells are cultured in interleukin-2 conditioned medium for several days for expansion. There is an option to expand CAR T cells further by co-incubation with irradiated, antigen-expressing feeder cell lines. The CAR T cells are ready to use after 10 (without feeder cell expansion) to 24 days (with feeder cell expansion). © 2020 The Authors. Basic Protocol: Generation of CAR T cells by lentiviral transduction.
Substances chimiques
Antibodies, Monoclonal
0
Biomarkers
0
Receptors, Antigen, T-Cell
0
Receptors, Chimeric Antigen
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e93Informations de copyright
© 2020 The Authors.
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