Understanding the freezing responses of T cells and other subsets of human peripheral blood mononuclear cells using DSMO-free cryoprotectants

Pi, Chia Hsing; Hornberger, Kathlyn; Dosa, Peter I.; Hubel, Allison

doi:10.1016/j.jcyt.2020.01.013

Cited by 19 publications

(16 citation statements)

References 38 publications

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“…This objective is especially important for adapting a cryopreservation method to different cell types or heterogeneous populations. Our recent study found that subsets of peripheral blood mononuclear cells (PBMCs) responded differently to MCOs made up of sugars, glycerol, and isoleucine 35 . The interactions between CPAs and cells may be cell‐type specific, indicating a need for custom CPA formulations for different cell types.…”

Section: Discussionmentioning

confidence: 99%

Natural deep eutectic systems for nature‐inspired cryopreservation of cells

Hornberger,

Li,

Duarte

et al. 2020

AIChE Journal

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Natural deep eutectic systems (NADES) are emerging as potential cryoprotective agents (CPA) for cell preservation. In this investigation, we develop an optimized CPA formulation using trehalose‐glycerol NADES (T:G) diluted in Normosol‐R and supplemented with isoleucine. Differential scanning calorimetry (DSC) is used to define the thermophysical properties of NADES‐based solutions, and Raman spectroscopy is used to characterize the effect of NADES on ice formation and hydrogen bonding. Jurkat cells are cryopreserved in each solution, and post‐thaw cell recovery, apoptosis, and growth are quantified. Raman spectra and heat maps show that NADES suppresses both ice formation and dehydration of the nonfrozen region. Supplementing NADES with isoleucine does not affect the solution's thermophysical properties but significantly improves the cells' survival and proliferation post‐thaw. The study indicates that thermophysical properties of CPA solutions alone cannot predict optimal cell survival, suggesting that stabilization of biological structures by CPAs may play a role in successful cryopreservation.

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Section: Discussionmentioning

confidence: 99%

Natural deep eutectic systems for nature‐inspired cryopreservation of cells

Hornberger,

Li,

Duarte

et al. 2020

AIChE Journal

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“…In general, cell samples can contain viable, apoptotic, dead cells or debris. In addition to Trypan blue, some other dyes are able to stain apoptotic or dead cells, which are subsequently analyzed with fluorescence microscopy 217,222 or flow cytometry 216,223 . Raman microscopy is another technique to discriminate viable from dead or apoptotic cells.…”

Section: Cell-based Medicinal Products (Cbmp)mentioning

confidence: 99%

“…The classical approach to identify cells is by labeling cell surface markers with fluorescent-dye conjugated antibodies followed by flow cytometry analysis. [215][216][217]223,225 In recent years, several label-free Raman spectroscopy approaches were developed for the phenotyping of immune cells. A line scan Raman method can be used to discriminate between immune cells 226 as well as a combination of single point Raman spectra and digital holographic microscopy 227 .…”

Section: Cell-based Medicinal Products (Cbmp)mentioning

confidence: 99%

Particles in Biopharmaceutical Formulations, Part 2: An Update on Analytical Techniques and Applications for Therapeutic Proteins, Viruses, Vaccines and Cells

Roesch

Zölls²,

Stadler³

et al. 2022

Journal of Pharmaceutical Sciences

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Particles in biopharmaceutical formulations remain a hot topic in drug product development. With new product classes emerging it is crucial to discriminate particulate active pharmaceutical ingredients from particulate impurities. Technical improvements, new analytical developments and emerging tools (e.g., machine learning tools) increase the amount of information generated for particles. For a proper interpretation and judgment of the generated data a thorough understanding of the measurement principle, suitable application fields and potential limitations and pitfalls is required. Our review provides a comprehensive overview of novel particle analysis techniques emerging in the last decade for particulate impurities in therapeutic protein formulations (protein-related, excipient-related and primary packaging material-related), as well as particulate biopharmaceutical formulations (virus particles, virus-like particles, lipid nanoparticles and cellbased medicinal products). In addition, we review the literature on applications, describe specific analytical approaches and illustrate advantages and drawbacks of currently available techniques for particulate biopharmaceutical formulations.

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“…200 to 1600 mOsm/kg) and the post-thaw recovery of the Jurkat cells. A recent paper from the same group evinced that sucrose could be substituted for trehalose at the same concentration and ratio to glycerol:isoleucine to maintain the post-thaw recovery of Jurkat cells and PBMCs near equivalent to 10% DMSO [ 107 ]. Substitution of sucrose for maltose was equally effective, although maltose is an interesting choice of disaccharide since it is a reducing sugar and risks the degradation of T cell surface proteins through the Maillard reaction, resulting in the formation of covalent adducts [ 108 ].…”

Section: Novel Excipients and Formulations Of T Cellsmentioning

confidence: 99%

Formulation Considerations for Autologous T Cell Drug Products

et al. 2021

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Genetically modified autologous T cells have become an established immunotherapy in the fight against cancer. The manufacture of chimeric antigen receptor (CAR) and αβ-T cell receptor (TCR) transduced T cells poses unique challenges, including the formulation, cryopreservation and fill–finish steps, which are the focus of this review. With an increasing number of marketing approvals for CAR-T cell therapies, comparison of their formulation design and presentation for administration can be made. These differences will be discussed alongside the emergence of automated formulation and fill-finish processes, the formulation design space, Monte Carlo simulation applied to risk analysis, primary container selection, freezing profiles and thaw and the use of dimethyl sulfoxide and alternative solvents/excipients as cryopreservation agents. The review will conclude with a discussion of the pharmaceutical solutions required to meet the simplification of manufacture and flexibility in dosage form for clinical treatment.

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Understanding the freezing responses of T cells and other subsets of human peripheral blood mononuclear cells using DSMO-free cryoprotectants

Cited by 19 publications

References 38 publications

Natural deep eutectic systems for nature‐inspired cryopreservation of cells

Natural deep eutectic systems for nature‐inspired cryopreservation of cells

Particles in Biopharmaceutical Formulations, Part 2: An Update on Analytical Techniques and Applications for Therapeutic Proteins, Viruses, Vaccines and Cells

Formulation Considerations for Autologous T Cell Drug Products

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