Comparison of chimeric antigen receptor-T cell-mediated cytotoxicity assays with suspension tumor cells using plate-based image cytometry method.

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  • Author(s): Sun YJ;Sun YJ; Chen YC; Chen YC; Hua WK; Hua WK; Wu SC; Wu SC; Chan LL; Chan LL
  • Source:
    Cytometry. Part A : the journal of the International Society for Analytical Cytology [Cytometry A] 2023 Jan; Vol. 103 (1), pp. 27-38. Date of Electronic Publication: 2022 Aug 03.
  • Publication Type:
    Journal Article
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Wiley-Liss Country of Publication: United States NLM ID: 101235694 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1552-4930 (Electronic) Linking ISSN: 15524922 NLM ISO Abbreviation: Cytometry A Subsets: MEDLINE
    • Publication Information:
      Original Publication: Hoboken, N.J. : Wiley-Liss, c2002-
    • Subject Terms:
    • Abstract:
      In the recent decade, chimeric antigen receptor (CAR)-T cell therapy has revolutionized strategies for cancer treatments due to its highly effective clinical efficacy and response for B cell malignancies. The success of CAR-T cell therapy has stimulated the increase in the research and development of various CAR constructs to target different tumor types. Therefore, a robust and efficient in vitro potency assay is needed to quickly identify potential CAR gene design from a library of construct candidates. Image cytometry methodologies have been utilized for various CAR-T cell-mediated cytotoxicity assay using different fluorescent labeling methods, mainly due to their ease-of-use, ability to capture cell images for verification, and higher throughput performance. In this work, we employed the Celigo Image Cytometer to evaluate and compare two CAR-T cell-mediated cytotoxicity assays using GFP-expressing or fluorescent dye-labeled myeloma and plasmacytoma cells. The GFP-based method demonstrated higher sensitivity in detecting CAR-T cell-mediated cytotoxicity when compared to the CMFDA/DAPI viability method. We have established the criteria and considerations for the selection of cytotoxicity assays that are fit-for-purpose to ensure the results produced are meaningful for the specific testing conditions.
      (© 2022 International Society for Advancement of Cytometry.)
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    • Contributed Indexing:
      Keywords: CAR-T cell-mediated cytotoxicity; Celigo; chimeric antigen receptor; immuno-oncology; plate-based image cytometry
    • Accession Number:
      0 (Receptors, Chimeric Antigen)
    • Publication Date:
      Date Created: 20220723 Date Completed: 20230119 Latest Revision: 20230205
    • Publication Date:
      20240105
    • Accession Number:
      10.1002/cyto.a.24673
    • Accession Number:
      35869932