Progress in Cryopreservation of Stem Cells and Immune Cells for Cytotherapy

Shu, Zhiquan; Heimfeld, Shelly; Huang, Zhongping; Liu, Carolyn; Gao, Dayong

doi:10.5772/60620

Cited by 6 publications

(3 citation statements)

References 84 publications

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“…4. Visually inspect the sample, and when only a small ice crystal remains visible, remove the sample and quickly begin any washing or post-thaw steps [23]. As a rule of the thumb, 2 mL cryovials containing 1 mL of liquid take~2 min to thaw from approximately À120 C in a 37 C water bath (see Note 19).…”

Section: Thawingmentioning

confidence: 99%

Freezing Technology: Control of Freezing, Thawing, and Ice Nucleation

Kilbride

Meneghel

2020

Methods in Molecular Biology

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From early dry-ice-based freezers and passive coolers, cryopreservation devices have come a long way. With increasing interest in the field of cryobiology from new scientific applications, the importance of reliable, traceable, and reproducible cold chain devices is sure to increase, ensuring more precise cryopreservation and enabling better post-thaw outcomes, both for the user and for biological samples. As with any cryopreservation process, it is important to optimize each part of the cold chain for each lab’s biological samples, cryocontainers used, and logistical restraints. In this chapter we describe how freezing technology can be used for cryopreservation of cells.

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Section: Thawingmentioning

confidence: 99%

Freezing Technology: Control of Freezing, Thawing, and Ice Nucleation

Kilbride

Meneghel

2020

Methods in Molecular Biology

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“…This was despite the longer time taken to complete thawing in the water-free thawing system when compared to the conventional water bath technique (means of 405 vs. 337 seconds respectively, p-value < 0.05). This result may appear unexpected as, across the broader field of cryopreservation, rapid thawing (at least as fast as can be achieved in a 37˚C water bath) is considered essential for good post-thaw recovery for a very wide range of cell types [26][27][28][29]. However, recent studies have shown that rapid thawing at this level is not required for somatic mammalian cells as long as the earlier cooling stage is appropriately controlled (as is the case with apheresis samples).…”

Section: Discussionmentioning

confidence: 99%

Automated dry thawing of cryopreserved haematopoietic cells is not adversely influenced by cryostorage time, patient age or gender

et al. 2020

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Cell therapies are becoming increasingly widely used, and their production and cryopreservation should take place under tightly controlled GMP conditions, with minimal batch-to-batch variation. One potential source of variation is in the thawing of cryopreserved samples, typically carried out in water baths. This study looks at an alternative, dry thawing, to minimise variability in the thawing of a cryopreserved cell therapy, and compares the cellular outcome on thaw. Factors such as storage time, patient age, and gender are considered in terms of cryopreservation and thawing outcomes. Cryopreserved leukapheresis samples from 41 donors, frozen by the same protocol and stored for up to 17 years, have been thawed using automated, water-free equipment and by conventional wet thawing using a water bath. Post-thaw viability, assessed by both trypan blue and flow cytometry, showed no significant differences between the techniques. Similarly, there was no negative effect of the duration of frozen storage, donor age at sample collection or donor gender on post-thaw viability using either thawing method. The implication of these results is that the cryopreservation protocol chosen initially remains robust and appropriate for use with a wide range of donors. The positive response of the samples to water-free thawing offers potential benefits for clinical situations by removing the subjective element inherent in water bath thawing and eliminating possible contamination issues.

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“…Studies have shown that preservation in −80 • C freezers provides long-term storage of tissues and cells from weeks to months, and is one of the most commonly used storage methods. The combination of slow freezing and fast thawing methods employed in this study is considered to be an optimal cryopreservation method, which induces less injury and damage to the cells and tissue (Shu et al 2015).…”

Section: Methodsmentioning

confidence: 99%

Monitoring the quality of frozen-thawed venous segments using bioimpedance spectroscopy

et al. 2020

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Objective: Storage at temperatures as low as −80 °C and below (cryopreservation) is considered a method for long-term preservation of cells and tissues, and especially blood vessel segments, which are to be used for clinical operations such as transplantation. However, the freezing and thawing processes themselves can induce injuries to the cells and tissue by damaging the structure and consequently functionality of the cryopreserved tissue. In addition, the level of damage is dependent on the rate of cooling and warming used during the freezing-thawing process. Current methods for monitoring the viability and integrity of cells and tissues after going through the freezing-thawing cycle are usually invasive and destructive to the cells and tissues. Therefore, employing monitoring methods which are not destructive to the cryopreserved tissues, such as bioimpedance measurement techniques, is necessary. In this study we aimed to design a bioimpedance measurement setup to detect changes in venous segments after freezing-thawing cycles in a noninvasive manner. Approach: A bioimpedance spectroscopy measurement technique with a two-electrode setup was employed to monitor ovine jugular vein segments after each cycle during a process of seven freezing-thawing cycles. Main results: The results demonstrated changes in the impedance spectra of the measured venous segments after each freezing-thawing cycle. Significance: This indicates that bioimpedance spectroscopy has the potential to be developed into a novel method for non-invasive and non-destructive monitoring of the viability of complex tissue after cryopreservation.

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Progress in Cryopreservation of Stem Cells and Immune Cells for Cytotherapy

Cited by 6 publications

References 84 publications

Freezing Technology: Control of Freezing, Thawing, and Ice Nucleation

Freezing Technology: Control of Freezing, Thawing, and Ice Nucleation

Automated dry thawing of cryopreserved haematopoietic cells is not adversely influenced by cryostorage time, patient age or gender

Monitoring the quality of frozen-thawed venous segments using bioimpedance spectroscopy

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