CAR-T Cell Manufacturing: Major Process Parameters and Next-generation Strategies

Overview

Journal J Exp Med

Specialties Allergy & Immunology
General Medicine

Date 2024 Jan 16

PMID 38226974

Authors

Melanie Ayala Ceja

Mobina Khericha

Caitlin M Harris

Cristina Puig-Saus

Yvonne Y Chen

Affiliations

Soon will be listed here.

Abstract

Chimeric antigen receptor (CAR)-T cell therapies have demonstrated strong curative potential and become a critical component in the array of B-cell malignancy treatments. Successful deployment of CAR-T cell therapies to treat hematologic and solid cancers, as well as other indications such as autoimmune diseases, is dependent on effective CAR-T cell manufacturing that impacts not only product safety and efficacy but also overall accessibility to patients in need. In this review, we discuss the major process parameters of autologous CAR-T cell manufacturing, as well as regulatory considerations and ongoing developments that will enable the next generation of CAR-T cell therapies.

Citing Articles

Enhanced conversion of T cells into CAR T cells by modulation of the MAPK/ERK pathway.

Adabi E, Charitidis F, Thalheimer F, Guaza-Lasheras M, Clarke C, Buchholz C Cell Rep Med. 2025; 6(2):101970.

PMID: 39938523 PMC: 11866553. DOI: 10.1016/j.xcrm.2025.101970.

Taming Variability in T-Cell Mechanosensing.

Schultheiss P, Pulkundwar A, Li W, Kam L Cells. 2025; 14(3).

PMID: 39936994 PMC: 11817355. DOI: 10.3390/cells14030203.

Challenges and overcoming strategies in CAR-T cell therapy for pediatric neuroblastoma.

Ying P, Tang Y World J Pediatr. 2025; 21(2):123-130.

PMID: 39900866 DOI: 10.1007/s12519-025-00876-9.

A Systematic Review on the Dual Role of Interleukin-1 in CAR T-Cell Therapy: Enhancer and Mitigator.

Maali A, Noei A, Feghhi-Najafabadi S, Sharifzadeh Z Iran Biomed J. 2025; 28(5 & 6):221-34.

PMID: 39891450 PMC: 11829154. DOI: 10.61186/ibj.4444.

Decoding the Implications of Zinc in the Development and Therapy of Leukemia.

Zhu B, Yang C, Hua S, Li K, Shang P, Li Z Adv Sci (Weinh). 2025; 12(9):e2412225.

PMID: 39887881 PMC: 11884550. DOI: 10.1002/advs.202412225.

References

Jing R, Scarfo I, Najia M, Lummertz da Rocha E, Han A, Sanborn M . EZH1 repression generates mature iPSC-derived CAR T cells with enhanced antitumor activity. Cell Stem Cell. 2022; 29(8):1181-1196.e6. PMC: 9386785. DOI: 10.1016/j.stem.2022.06.014. View

Levine B, Miskin J, Wonnacott K, Keir C . Global Manufacturing of CAR T Cell Therapy. Mol Ther Methods Clin Dev. 2017; 4:92-101. PMC: 5363291. DOI: 10.1016/j.omtm.2016.12.006. View

Ruella M, Xu J, Barrett D, Fraietta J, Reich T, Ambrose D . Induction of resistance to chimeric antigen receptor T cell therapy by transduction of a single leukemic B cell. Nat Med. 2018; 24(10):1499-1503. PMC: 6511988. DOI: 10.1038/s41591-018-0201-9. View

Maude S, Laetsch T, Buechner J, Rives S, Boyer M, Bittencourt H . Tisagenlecleucel in Children and Young Adults with B-Cell Lymphoblastic Leukemia. N Engl J Med. 2018; 378(5):439-448. PMC: 5996391. DOI: 10.1056/NEJMoa1709866. View

Berdeja J, Madduri D, Usmani S, Jakubowiak A, Agha M, Cohen A . Ciltacabtagene autoleucel, a B-cell maturation antigen-directed chimeric antigen receptor T-cell therapy in patients with relapsed or refractory multiple myeloma (CARTITUDE-1): a phase 1b/2 open-label study. Lancet. 2021; 398(10297):314-324. DOI: 10.1016/S0140-6736(21)00933-8. View

Trainor N, Purpura K, Middleton K, Fargo K, Hails L, Vicentini-Hogan M . Automated production of gene-modified chimeric antigen receptor T cells using the Cocoon Platform. Cytotherapy. 2023; 25(12):1349-1360. DOI: 10.1016/j.jcyt.2023.07.012. View

Bos R, Sherman L . CD4+ T-cell help in the tumor milieu is required for recruitment and cytolytic function of CD8+ T lymphocytes. Cancer Res. 2010; 70(21):8368-77. PMC: 2970736. DOI: 10.1158/0008-5472.CAN-10-1322. View

Long A, Haso W, Shern J, Wanhainen K, Murgai M, Ingaramo M . 4-1BB costimulation ameliorates T cell exhaustion induced by tonic signaling of chimeric antigen receptors. Nat Med. 2015; 21(6):581-90. PMC: 4458184. DOI: 10.1038/nm.3838. View

Deng Q, Han G, Puebla-Osorio N, Ma M, Strati P, Chasen B . Characteristics of anti-CD19 CAR T cell infusion products associated with efficacy and toxicity in patients with large B cell lymphomas. Nat Med. 2020; 26(12):1878-1887. PMC: 8446909. DOI: 10.1038/s41591-020-1061-7. View

10.

Dias J, Cadinanos-Garai A, Roddie C . Release Assays and Potency Assays for CAR T-Cell Interventions. Adv Exp Med Biol. 2023; 1420:117-137. DOI: 10.1007/978-3-031-30040-0_8. View

11.

Teoh J, Brown L . Developing lisocabtagene maraleucel chimeric antigen receptor T-cell manufacturing for improved process, product quality and consistency across CD19 hematologic indications. Cytotherapy. 2022; 24(9):962-973. DOI: 10.1016/j.jcyt.2022.03.013. View

12.

Larson S, Walthers C, Ji B, Ghafouri S, Naparstek J, Trent J . CD19/CD20 Bispecific Chimeric Antigen Receptor (CAR) in Naive/Memory T Cells for the Treatment of Relapsed or Refractory Non-Hodgkin Lymphoma. Cancer Discov. 2022; 13(3):580-597. PMC: 9992104. DOI: 10.1158/2159-8290.CD-22-0964. View

13.

Wang X, Borquez-Ojeda O, Stefanski J, Du F, Qu J, Chaudhari J . Depletion of high-content CD14 cells from apheresis products is critical for successful transduction and expansion of CAR T cells during large-scale cGMP manufacturing. Mol Ther Methods Clin Dev. 2021; 22:377-387. PMC: 8411225. DOI: 10.1016/j.omtm.2021.06.014. View

14.

Depil S, Duchateau P, Grupp S, Mufti G, Poirot L . 'Off-the-shelf' allogeneic CAR T cells: development and challenges. Nat Rev Drug Discov. 2020; 19(3):185-199. DOI: 10.1038/s41573-019-0051-2. View

15.

Scholler J, Brady T, Binder-Scholl G, Hwang W, Plesa G, Hege K . Decade-long safety and function of retroviral-modified chimeric antigen receptor T cells. Sci Transl Med. 2012; 4(132):132ra53. PMC: 4368443. DOI: 10.1126/scitranslmed.3003761. View

16.

Good Z, Spiegel J, Sahaf B, Malipatlolla M, Ehlinger Z, Kurra S . Post-infusion CAR T cells identify patients resistant to CD19-CAR therapy. Nat Med. 2022; 28(9):1860-1871. PMC: 10917089. DOI: 10.1038/s41591-022-01960-7. View

17.

Casati A, Varghaei-Nahvi A, Feldman S, Assenmacher M, Rosenberg S, Dudley M . Clinical-scale selection and viral transduction of human naïve and central memory CD8+ T cells for adoptive cell therapy of cancer patients. Cancer Immunol Immunother. 2013; 62(10):1563-73. PMC: 6348480. DOI: 10.1007/s00262-013-1459-x. View

18.

Bhaskar S, Dholaria B, Sengsayadeth S, Savani B, Oluwole O . Role of bridging therapy during chimeric antigen receptor T cell therapy. EJHaem. 2022; 3(Suppl 1):39-45. PMC: 9175845. DOI: 10.1002/jha2.335. View

19.

Jain N, Zhao Z, Feucht J, Koche R, Iyer A, Dobrin A . TET2 guards against unchecked BATF3-induced CAR T cell expansion. Nature. 2023; 615(7951):315-322. PMC: 10511001. DOI: 10.1038/s41586-022-05692-z. View

20.

Chong E, Levine B, Grupp S, Davis M, Siegel D, Maude S . CAR T cell viability release testing and clinical outcomes: is there a lower limit?. Blood. 2019; 134(21):1873-1875. PMC: 6872962. DOI: 10.1182/blood.2019002258. View