![]() ![]() Manufacturing of clinical-grade NKG2D CAR memory T cells using CliniMACS Prodigy is feasible and reproducible, widening clinical application of CAR T cell therapies.ĬliniMACS prodigy NKG2D CAR automated production clinical-grade large-scale memory T cells.Ĭopyright © 2019 Fernández, Fernández, Mirones, Escudero, Cardoso, Vela, Lanzarot, de Paz, Leivas, Gallardo, Marcos, Romero, Martínez-López and Pérez-Martínez. ![]() CAR T cell final products met release criteria, except for one showing myc overexpression and another with viral copy number higher than five. The manufacturing protocol here described achieved large numbers of viable NKG2D CAR memory T cells with elevated levels of NKG2D CAR expression and highly cytotoxic against Jurkat and 531MII tumor target cells. Final cell products were analyzed to comply with the specifications derived from the quality and complementary controls carried out in accordance with the instructions of the Spanish Regulatory Agency of Medicines and Medical Devices (AEMPS) for the manufacture of investigational advanced therapy medicinal products (ATMPs). NKG2D CAR T cells expanded between 10 and 13 days. Then, we used NKG2D-CD8TM-4-1BB-CD3ζ lentiviral vector for cell transduction (MOI = 2). A total of 10 8 CD45RA - cells were cultured in TexMACS media supplemented with 100 IU/mL IL-2 and activated at day 0 with T Cell TransAct. CD45RA + fraction was depleted from healthy donors' non-mobilized apheresis using CliniMACS CD45RA Reagent and CliniMACS Plus device. Bioz Stars score: 80/100, based on 1 PubMed citations. In this study, we developed a protocol to obtain large-scale NKG2D CAR memory T cells for clinical use by using CliniMACS Prodigy, an automated closed system compliant with Good Manufacturing Practice (GMP) guidelines. Miltenyi Biotecgmp grade texmacs Gmp Grade Texmacs, supplied by Miltenyi Biotec, used in various techniques. To avoid such adverse effects, we used CD45RA - memory T cells, a T-cell subset with less alloreactivity, as effector cells to express NKG2D CAR. The use of allogeneic T cells for CAR therapy could be an attractive alternative, although undesirable graft vs. Patient-derived CAR T cells show limitations including inability to manufacture CAR T cells from the patients' own T cells, disease progression, and death prior to return of engineered cells. A second-generation NKG2D CAR was generated by fusing full-length human NKG2D to 4-1BB costimulatory molecule and CD3ζ signaling domain. NKG2D chimeric antigen receptor (CAR) T cells have shown potent anticancer effects against different cancer types. Natural killer group 2D (NKG2D) is a natural killer (NK) cell-activating receptor that recognizes different stress-induced ligands that are overexpressed in a variety of childhood and adult tumors.
0 Comments
Leave a Reply. |