Cryopreservation adds flexibility to timelines when working with cells. In the case of cellular therapies, that can mean initial flexibility to schedule patient cell collection, transport cells to the manufacturing facility and initiate manufacturing. Cryopreservation of the final cell therapy product adds flexibility to be able to schedule the return of cells to the patient care site and schedule the patient to receive therapeutic cells. This article by Brudno et al. recently added to BioLifes’ Evidence Library provides a snapshot of CAR T cell preparation for use in a clinical trial. The CAR T cells were cryopreserved for storage in vapor-phase liquid nitrogen. Brudno, J. N., Lam, N., Vanasse, D., Shen, Y., Rose, J. J., Rossi, J., Xue, A., Bot, A., Scholler, N., Mikkilineni, L., Roschewski, M., Dean, R., Cachau, R., Youkharibache, P., Patel, R., Hansen, B., Stroncek, D. F., Rosenberg, S. A., Gress, R. E., Kochenderfer, J. N. Safety and feasibility of anti-CD19 CAR T cells with fully human binding domains in patients with B-cell lymphoma. Nature Medicine. (2020). doi: 10.1038/s41591-019-0737-3 Brudno et al. reported results from their first-in-human trial of anti-CD19 CAR Hu19-cd838? featuring fully human binding domains used to treat 20 patients with B-cell lymphoma (NCT02659943). The authors also compared results from patients treated with Hu19-cd828? CAR T cells to results from patients treated with anti-CD19 CAR FMC63-28? T cells, in commercial development at the time as axicabtagene ciloleucel, in clinical trial NCT00924326. Fifteen study patients in clinical trial NCT00924326 received cryopreserved FMC63-28? T cells. According to the authors, FMC63-28? T cells were manufactured, washed, and prepared for cryopreservation as follows: “On the day of cryopreservation, the [FMC63-28?] cells were washed with 0.9% saline with the Sepax II. The FMC63-28? T-cell product was formulated in CS250 cryostorage bags (OriGen Biomedical) at the target dose. T cells were suspended in a solution containing 0.9% saline plus 5% HAS, and a 1:1 volume of Cryostor 10 (BioLife Solutions) was added. The cells were frozen in a controlled-rate freezer (Planer) and stored in vapor-phase liquid nitrogen. On the day of infusion, the frozen cells were thawed at 37℃ and infused into patients within 2 h.” The purpose for cryopreserving cells in the following article by Lam et al. was to freeze transduced patient T cells for later use as a source of antigen-presenting cells to stimulate autologous peripheral blood mononuclear cells in culture. Lam, N., Trinklein, N., Buelow, B., Patterson, G., Ojha, N., Kochenderfer, J. Anti-BCMA chimeric antigen receptors with fully human heavy-chain-only antigen recognition domains. Nature Communications. 2020;11(1). doi: 10.1038/s41467-019-14119-9 The authors investigated a murine scFv binding domain as a potential source of anti-CAR immune responses in patients receiving CAR T cells in clinical trials. A non-signaling 11D5-3-NS chimeric antigen receptor analogous to 11D5-3-CD828Z, minus the signaling domain, was designed to be delivered via gamma-retroviral vector to T cells from patients that had previously received 11D5-3-CD828Z CAR T cell therapy. A separate set of patient T cells were transduced with a gamma retrovirus encoding human full-length low-affinity NGFR to be used as a negative control. Both sets of T cells were cryopreserved in CryoStor CS10 for later use. Thawed and irradiated, the cells served as antigen-presenting cells, cultured with autologous, expanded peripheral blood mononuclear cells (PBMCs) from patients who had received 11D5-3-CD828Z CAR T cell therapy. I already freeze my cells in CryoStor®. What else can BioLife do for me? BioLife Solutions has expanded our portfolio of cell and gene therapy tools originating with best-in-class CryoStor and HypoThermosol® biopreservation media, by acquiring Astero Bio Corporation, Custom Biogenic Systems and SAVSU Technologies. Meet ThawSTAR CB, the newest member of first-of-its-kind automated ThawSTAR technology for water-free thawing of cells frozen in 25 mL to 1000 mL cryobags. View the ThawSTAR CB video here! The ThawSTAR product lineup includes ThawSTAR systems for cells frozen in vials: CFT1.5 (1.5 mL cryovials), CFT2 (1.8 – 2.0 mL cryovials), and AT6 (6.0 mL AT-Closed Vial®). How do you handle temperature-sensitive frozen biosamples? Check out the BioT line of transporters (ULT, LN2) and the ULT Mobile Workstation. Need a controlled rate freezer? Consider Custom Biogenic Systems’ Series 2101 Controlled Rate Freezers. For storing cells frozen in vapor-phase liquid nitrogen, CBS’ line of Isothermal Liquid Nitrogen Freezers (patented) maximizes storage space while minimizing chances of cross-contamination. Ship your cell and gene therapies in evo® DV-4 and DV-10 liquid nitrogen dry vapor smart shippers developed by SAVSU Technologies. The evo system features the most advanced smart containers and Software-as-a-Service technologies that enable specialty couriers to offer a higher level of service to you and those you serve. Need more information? Visit our website at www.biolifesolutions.com. Search our Evidence Library. Ask the Scientists. To place an order or request more information, call us today! 1.866.6543 (toll-free in North America) or direct: 1.425.402.1400.