Impact of cryopreservation on CAR T production and clinical response
Open accessAbstract
Adoptive cell therapy with chimeric antigen receptor (CAR) T cells has become an efficient treatment option for patients with hematological malignancies. FDA approved CAR T products are manufactured in centralized facilities from fresh or frozen leukapheresis and the cryopreserved CAR T infusion product is shipped back to the patient. An increasing number of clinical centers produce CAR T cells on-site, which enables the use of fresh and cryopreserved PBMCs and CAR T cells. Here we determined the effect of cryopreservation on PBMCs and CD19 CAR T cells in a cohort of 118 patients treated with fresh CAR T cells and in several patients head-to-head. Cryopreserved PBMCs, obtained from leukapheresis products, contained less erythrocytes and T cells, but were sufficient to produce CAR T cells for therapy. There was no correlation between the recovery of PBMCs and the transduction efficacy, the number of CAR T cells obtained by the end of the manufacturing process, the in vitro reactivity, or the response rate to CAR T therapy. We could show that CAR T cells cryopreserved during the manufacturing process, stored and resumed expansion at a later time point, yielded sufficient cell numbers for treatment and led to complete remissions. Phenotype analysis including T cell subtypes, chemokine receptor and co-inhibitory/stimulatory molecules, revealed that fresh CAR T cells expressed significantly more TIM-3 and contained less effector T cells in comparison to their frozen counterparts. In addition, fresh CAR T infusion products demonstrated increased in vitro anti-tumor reactivity, however cryopreserved CAR T cells still showed high anti-tumor potency and specificity. The recovery of cryopreserved CAR T cells was similar in responding and non-responding patients. Although fresh CAR T infusion products exhibit higher anti-tumor reactivity, the use of frozen PBMCs as staring material and frozen CAR T infusion products seems a viable option, as frozen products still exhibit high in vitro potency and cryopreservation did not seem to affect the clinical outcome.
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Fresh CAR-T cells were shown earlier to have higher in vitro antitumor activity and lesser microcellular damage than frozen CAR-T cells.
“POC manufacturing enables the delivery of fresh CAR-T cells, which were shown earlier to have higher in vitro antitumor activity and lesser microcellular damage than the frozen,4,5”
From Point-of-care manufacturing of anti-CD19 CAR-T cells using a closed production platform: Experiences of an academic in Thailand (2024) · cites this paper as [4]
The Discussion reports higher in vitro anti-tumor reactivity in fresh CAR T infusion products than in cryopreserved products. The Results describe differences in cell phenotype, but do not establish that fresh CAR T cells have less microcellular damage. No page numbers are available in our copy. (LLM, conf=0.86)
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Miller, B. C., Sen, D. R., Al Abosy, R., Bi, K., Virkud, Y. V., LaFleur, M. W., Yates, K. B., Lako, A., Felt, K., Naik, G. S., et al. (2019) · Nature Immunology · doi:10.1038/s41590-019-0312-6 · Cited by 2,087 · see citing works
Effect of cryopreservation on autologous chimeric antigen receptor T cell characteristics
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Luanpitpong, S., Klaihmon, P., Janan, M., Kungwankiattichai, S., Owattanapanich, W., Kunacheewa, C., Chanthateyanonth, S., Donsakul, N., U-pratya, Y., Warindpong, T., et al. (2024) · Molecular Therapy: Oncology · doi:10.1016/j.omton.2024.200889 · Cited by 17 · see citing works
“POC manufacturing enables the delivery of fresh CAR-T cells, which were shown earlier to have higher in vitro antitumor activity and lesser microcellular damage than the frozen, 4,5 although whether fresh products would provide better clinical outcomes remains to be determined.”
- Authors
- Brezinger-Dayan, K., Itzhaki, O., Melnichenko, J., Kubi, A., Zeltzer, L.-A., Jacoby, E., Avigdor, A., Shapira Frommer, R., & Besser, M. J.
- Affiliations
- Sheba Medical Center (IL)
- Tel Aviv University (IL)
- Corresponding author
- Itzhaki, Orit
- Besser, Michal J.
- Venue
- Frontiers in Oncology
- Publisher
- Frontiers Media SA
- Published
- 2022-10-06
- Volume
- 12
- Pages
- 1024362-1024362
- ISSN
- 2234-943X (ISSN-L)
- PMC
- PMC9582437
- OpenAlex
- W4302303770
- Cited by
- 44
- References
- 2
- Citations by year
- 2026: 10 · 2025: 20 · 2024: 10 · 2023: 3 · 2022: 1
- FWCI
- 3.56
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Frontiers in Oncology (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Topics
- Health Sciences › Medicine › Oncology › CAR-T cell therapy research
- Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Keywords
- CAR T-cell therapy, cryopreservation, CD19 CAR T cells, hematologic malignancies, leukapheresis, transduction efficiency, TIM-3, T cell exhaustion, clinical response, complete remission
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