Preservation of cell-based immunotherapies for clinical trials

Rui Li, Rachel Johnson, Guanglin Yu, David H. McKenna, Allison Hubel

Research output: Contribution to journalReview articlepeer-review

56 Scopus citations


In the unique supply chain of cellular therapies, preservation is important to keep the cell product viable. Many factors in cryopreservation affect the outcome of a cell therapy: (i) formulation and introduction of a freezing medium, (ii) cooling rate, (iii) storage conditions, (iv) thawing conditions and (v) post-thaw processing. This article surveys clinical trials of cellular immunotherapy that used cryopreserved regulatory, chimeric antigen receptor or gamma delta T cells, dendritic cells or natural killer (NK) cells. Several observations are summarized from the given information. The aforementioned cell types have been similarly frozen in media containing 5–10% dimethyl sulfoxide (DMSO) with plasma, serum or human serum albumin. Two common freezing methods are an insulated freezing container such as Nalgene Mr. Frosty and a controlled-rate freezer at a cooling rate of -1°C/min. Water baths at approximately 37°C have been commonly used for thawing. Post-thaw processing of cryopreserved cells varied greatly: some studies infused the cells immediately upon thawing; some diluted the cells in a carrier solution of varying formulation before infusion; some washed cells to remove cryoprotective agents; and others re-cultured cells to recover cell viability or functionality lost due to cryopreservation. Emerging approaches to preserving cellular immunotherapies are also described. DMSO-free formulations of the freezing media have demonstrated improved preservation of cell viability in T lymphocytes and of cytotoxic function in natural killer cells. Saccharides are a common type of molecule used as an alternative cryoprotective agent to DMSO. Improving methods of preservation will be critical to growth in the clinical use of cellular immunotherapies.

Original languageEnglish (US)
Pages (from-to)943-957
Number of pages15
Issue number9
StatePublished - Sep 2019

Bibliographical note

Funding Information:
This work was supported by the National Institute of Health ( R01EB023880 ).

Publisher Copyright:
© 2019 International Society for Cell and Gene Therapy


  • T cells
  • cell therapies
  • cryopreservation
  • dendritic cells
  • natural killer cells


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