Preclinical testing of CAR T cells in zebrafish xenografts

Methods Cell Biol. 2022;167:133-147. doi: 10.1016/bs.mcb.2021.07.002. Epub 2021 Aug 20.


Engineered chimeric antigen receptor T cells (CAR T cells) have emerged as a promising immunotherapy for cancer and have proven to be effective for B cell malignancies. Currently, great efforts are undertaken to expand the application of CAR T cells to other cancer entities, to increase the efficacy of CAR T cell-mediated killing of cancer cells and to reduce possible side effects of CAR T cell therapy. This creates a need for preclinical models to test the many emerging novel CAR designs. Traditionally, mouse xenograft models are applied to investigate the efficacy of CAR T cells in vivo. Here, we describe a complementing xenograft protocol for testing CAR T cells against human leukemia cells in zebrafish embryos. The embryonic zebrafish xenograft promises to be a fast and cost-efficient model and particularly offers live imaging opportunities of CAR T cell distribution and killing of cancer cells in vivo.

Keywords: CAR T cells; Cancer; Leukemia; Preclinical testing; Xenotransplantation; Zebrafish.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Heterografts
  • Humans
  • Immunotherapy, Adoptive / methods
  • Mice
  • Receptors, Antigen, T-Cell / genetics
  • Receptors, Chimeric Antigen* / genetics
  • Receptors, Chimeric Antigen* / therapeutic use
  • T-Lymphocytes
  • Zebrafish*


  • Receptors, Antigen, T-Cell
  • Receptors, Chimeric Antigen