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Cryopreserved leukapheresis enables scalable and distributed CAR-T manufacturing: a multi-platform comparative study.

Menglin Ren1,2, Xiaodie Wang3, Jiangrui Chen2

  • 1Shanghai Mengchao Cancer Hospital, Shanghai University, 118 Qianyang Road, Shanghai, 201805, China.

Scientific Reports
|August 14, 2025
PubMed
Summary

Cryopreserved leukapheresis offers a scalable, standardized source for Chimeric antigen receptor T (CAR-T) cell therapy manufacturing. This study validates its quality and functionality, ensuring consistent CAR-T production and supply chain resilience.

Keywords:
CAR-TCryopreserved leukapheresisFast CAR-TPBMCs

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Area of Science:

  • Cellular immunotherapy
  • Bioprocessing and manufacturing
  • Hematology

Background:

  • Cryopreserved leukapheresis is an underutilized but scalable source for Chimeric antigen receptor T (CAR-T) manufacturing.
  • Current limitations include a lack of standardized preparation and low automation levels.

Purpose of the Study:

  • To develop and validate a standardized, automated cryopreservation process for leukapheresis.
  • To systematically evaluate the quality, functionality, and platform compatibility of cryopreserved leukapheresis for CAR-T production.

Main Methods:

  • Optimized centrifugation, CS10 proportion, and cryopreservation procedures within a closed automated system.
  • Evaluated post-thaw viability, cell recovery, phenotypic profiles, and functional CAR-T attributes.
  • Assessed performance across non-viral, lentiviral, and Fast CAR-T platforms compared to fresh leukapheresis.

Main Results:

  • Established a standardized cryopreserved leukapheresis process yielding ≥90% post-thaw viability.
  • Cryopreserved leukapheresis showed comparable recovery and phenotype to peripheral blood mononuclear cells (PBMCs), with a higher lymphocyte proportion.
  • Demonstrated comparable CAR-T cell viability, expansion, phenotype, CAR+ cell proportion, and cytotoxicity across different CAR-T platforms.

Conclusions:

  • Cryopreserved leukapheresis is validated as a universal raw material for CAR-T manufacturing, preserving critical quality attributes and ensuring consistency.
  • Standardization and automation enhance its utility, improving supply chain resilience by decoupling from fresh material logistics.
  • Further large-scale clinical validation is recommended to solidify its role in CAR-T production.