Cryopreserved anti-CD22 and bispecific anti-CD19/22 CAR T cells are as effective as freshly infused cells

Manufacture of CD22 CAR T cells following positive versus negative selection results in distinct cytokine secretion profiles and γδ T cell output

“…Of note, cryopreserved MNC products have been found to be equivalent to their freshly isolated counterparts as regards expansion, CAR transduction, and patient responses. 42 , 43 Positive T cell selection was performed using CliniMACS Plus (Miltenyi Biotec) with anti-CD4 and anti-CD8 GMP magnetic selection beads (Miltenyi Biotec) for a combined CD4/CD8 selection. Briefly, thawed cells were washed twice with PBS/EDTA + 0.5% HSA to remove platelets, blocked with 1.5 mg/mL intravenous immunoglobulin (Baxter Healthcare) for 5 min and then labeled with magnetic capture beads for 30 min at room temperature.…”

Programming CAR T Cell Tumor Recognition: Tuned Antigen Sensing and Logic Gating

“…Cryopreservation enables the storage and distribution of drug products. The efficacy of cryopreserved CAR T cells was demonstrated to be comparable to fresh CAR T cells upon measuring in vivo expansion, persistence, incidence of toxicities, and disease response ( 136 ). Based on this study and data from two clinical trials (NCT02315612 and NCT03448393), CAR T cells can be cryopreserved without altering their functionality, providing greater flexibility for scheduling infusions and delivery to CAR T cell administration sites.…”

Post-thaw application of ROCK-inhibitors increases cryopreserved T-cell yield

“…44 Although results vary along studies, their cryopreservation is usually supported, as evidenced by the FDA approval of cryopreserved CAR-T products, which allow their centralised manufacturing. 45–48…”

“…44 Although results vary along studies, their cryopreservation is usually supported, as evidenced by the FDA approval of cryopreserved CAR-T products, which allow their centralised manufacturing. [45][46][47][48] Biochemical pathways dysregulated after cryopreservation will differ between cell types, therefore it is important to approach this in a "cell-type dependent" manner. 49 The literature suggests that T-cells are affected by apoptosis after cryopreservation, with around 40% of cells undergoing apoptosis 8 h post-thaw, followed by extensive cell death as shown by Sarkar et al 50 In Jurkat cells (a model T-cell line) inhibition of apoptosis using a pan-caspase inhibitor improved cell recovery and survival.…”