An update on methods for cryopreservation and thawing of hemopoietic stem cells

Lecchi, Lucilla; Giovanelli, S.; Gagliardi, Barbara Sara Alessandra; Pezzali, Ilaria; Ratti, Ilaria; Marconi, Maurizio

doi:10.1016/j.transci.2016.05.009

Cited by 46 publications

(41 citation statements)

References 73 publications

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“…Conventional, centrifuge-based methods for washing are time-consuming, result in cell losses which can prolong platelet engraftment, and pose a risk of cell contamination [27, 49, 58]. To alleviate these issues, alternatives to washing include freezing cells in a lower concentration of DMSO or diluting the thawed product before infusion.…”

Section: New Technologymentioning

confidence: 99%

Cryopreservation of Hematopoietic Stem Cells: Emerging Assays, Cryoprotectant Agents, and Technology to Improve Outcomes

Hornberger

McKenna

et al. 2019

Transfus Med Hemother

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Hematopoietic stem cell (HSC) therapy is widely used to treat a growing number of hematological and non-hematological diseases. Cryopreservation of HSCs allows for cells to be transported from the site of processing to the site of clinical use, creates a larger window of time in which cells can be administered to patients, and allows sufficient time for quality control and regulatory testing. Currently, HSCs and other cell therapies conform to the same cryopreservation techniques as cells used for research purposes: cells are cryopreserved in dimethyl sulfoxide (DMSO) at a slow cooling rate. As a result, HSC therapy can result in numerous adverse symptoms in patients due to the infusion of DMSO. Efforts are being made to improve the cryopreservation of HSCs for clinical use. This review discusses advances in the cryopreservation of HSCs from 2007 to the present. The preclinical development of new cryoprotectants and new technology to eliminate cryoprotectants after thawing are discussed in detail. Additional cryopreservation considerations are included, such as cooling rate, storage temperature, and cell concentration. Preclinical cell assessment and quality control are discussed, as well as clinical studies from the past decade that focus on new cryopreservation protocols to improve patient outcomes.

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Section: New Technologymentioning

confidence: 99%

Cryopreservation of Hematopoietic Stem Cells: Emerging Assays, Cryoprotectant Agents, and Technology to Improve Outcomes

Hornberger

McKenna

et al. 2019

Transfus Med Hemother

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“…Thawing may be carried out in the laboratory or at the bedside, with or without a washing step. Although washing procedures may in theory reduce the risk of complications due to remaining DMSO or cellular debris in the thawed product, this should be weighed against potential time delays which may impair cell quality and patient outcome [10]. …”

Section: Discussionmentioning

confidence: 99%

“…On the other hand, omitting the washing step may expose vulnerable stem cells to higher concentrations of cell debris and DMSO that can theoretically cause cellular injury and adverse events [3]. Washing the product in the laboratory after thawing and prior to infusion allows the removal of DMSO and cell fragments but may cause cell loss and requires more hands-on time and staff training [10]. …”

Section: Discussionmentioning

confidence: 99%

To Wash or Not to Wash? Comparison of Patient Outcome after Infusion of Cryopreserved Autologous Hematopoietic Stem Cells before and after the Replacement of Manual Washing by Bedside Thawing

et al. 2018

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Background: Prior to infusion, cryopreserved autologous peripheral blood stem cell (auto-PBSC) grafts can either be thawed at the bedside or thawed and washed at the laboratory. At our center, manual washing of grafts prior to infusion was discontinued in April 2012 and bedside thawing was implemented. Methods: This study compares the outcomes of two patient groups who received auto-PBSC either after post-thaw washing (n = 84) or bedside thawing (n = 83). Results: No life-threatening infusion-related side effects were reported in either group. There was no significant difference in the mean CD34+ cells/kg dose of infused auto-PBSC in the two groups (p = 0.41), nor in the number of days to neutrophils > 0.5 × 10⁹/L (p = 0.14), days to platelets > 20 × 10⁹/L (p = 0.64), or days to platelets > 50 × 10⁹/L (p = 0.62) after transplant. There was also no difference in the number of days on total parenteral nutrition (p = 0.69), days on G-CSF therapy (p = 0.48), or days with fever (p = 0.73). Finally, there was no significant difference in the number of red cell units transfused (p = 0.32), or platelet units transfused (p = 0.94) after the transplant. One-hundred-day mortality was identical in the two groups (2.4%). Conclusion: Both thawing procedures are safe and result in acceptable engraftment and patient outcomes.

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“…From the laboratory, to cryopreserved tissue allografts in theatre and point-of-care thawing of haematopoietic stem cells at the bedside [72], the almost universal use of this technology has led to most cryopreservation protocols being optimised against the back-drop of a warming rate achieved through its use. However, the use of such a method within a GMP environment raises issue of potential contamination and logistical problems in maintaining sterility.…”

Section: Thawing and Elution Of Cpamentioning

confidence: 99%

Technical Considerations in the Freezing, Low-Temperature Storage and Thawing of Stem Cells for Cellular Therapies

Hunt¹

2019

Transfus Med Hemother

105

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The commercial and clinical development of cellular therapy products will invariably require cryopreservation and frozen storage of cellular starting materials, intermediates and/or final product. Optimising cryopreservation is as important as optimisation of the cell culture process in obtaining maximum yield and a consistent end-product. Suboptimal cryopreservation can lead not only to batch-to-batch variation, lowered cellular functionality and reduced cell yield, but also to the potential selection of subpopulations with genetic or epigenetic characteristics divergent from the original cell line. Regulatory requirements also impact on cryopreservation as these will require a robust and reproducible approach to the freezing, storage and thawing of the product. This requires attention to all aspects of the application of low temperatures: from the choice of freezing container and cryoprotectant, the cooling rate employed and its mode of delivery, the correct handling of the frozen material during storage and transportation, to the eventual thawing of the product by the end-user. Each of these influences all of the others to a greater or lesser extent and none should be ignored. This paper seeks to provide practical insights and alternative solutions to the technical challenges faced during cryopreservation of cells for use in cellular therapies.

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An update on methods for cryopreservation and thawing of hemopoietic stem cells

Cited by 46 publications

References 73 publications

Cryopreservation of Hematopoietic Stem Cells: Emerging Assays, Cryoprotectant Agents, and Technology to Improve Outcomes

Cryopreservation of Hematopoietic Stem Cells: Emerging Assays, Cryoprotectant Agents, and Technology to Improve Outcomes

To Wash or Not to Wash? Comparison of Patient Outcome after Infusion of Cryopreserved Autologous Hematopoietic Stem Cells before and after the Replacement of Manual Washing by Bedside Thawing

Technical Considerations in the Freezing, Low-Temperature Storage and Thawing of Stem Cells for Cellular Therapies

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