Manufacture of B7-H3-haCAR-T cells
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(a) Scheme of B7-H3-haCAR which comprises of anti-B7-H3 scFv, the hinge and transmenbrane (TM) region of human CD8α, CD28 and 4-1BB signaling domain, human CD3ζ chain, IRES and ZsGreen. (b) and (c) The expression effectiveness of haCAR was detected through (b) fluorescent microscope and (c) flow cytometric assay of ZsGreen. Flow cytometry detected 25.2% of total T cells expressing ZsGreen. (d) Immunoblot analysis of cell lysates from both untransduced and CAR transduced T lymphocytes. (e) and (f) Phenotypic categories of NT and haCAR-T cells were determined by flow cytometric analysis after 14 days of lentiviral transduction
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