Improved cryopreservation yield of pancreatic islets using combination of lower dose permeable cryoprotective agents

Kojayan, Greg Garo; Whaley, David; Alexander, Michael P.; Rodriguez, Samuel; Lee, Sarah; Lakey, Jonathan R. T.

doi:10.1016/j.cryobiol.2019.04.004

Cited by 15 publications

(10 citation statements)

References 21 publications

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“…This has the benefit of a reduction in PA-induced toxicity and an increase in cellular viability and yields post-thaw. Another method that showed reduced CPA induced toxicity was demonstrated by Kojayan et al in which multi-molar combinations of reduced concentrations of EG, and DMSO were used to cryopreserve both human and murine islet cells 21 . They proposed that the reduction in adverse effects was likely due to reduced exposure to each individual CPA even though overall osmolality of the different combinations was comparable to a standard 2 M DMSO-only solution 21 .…”

Section: Introductionmentioning

confidence: 99%

“…Another method that showed reduced CPA induced toxicity was demonstrated by Kojayan et al in which multi-molar combinations of reduced concentrations of EG, and DMSO were used to cryopreserve both human and murine islet cells 21 . They proposed that the reduction in adverse effects was likely due to reduced exposure to each individual CPA even though overall osmolality of the different combinations was comparable to a standard 2 M DMSO-only solution 21 . Other studies have shown combinations of sucrose, propanediol, and DMSO to be effective at supporting post-warming survival of porcine blastocysts 15 .…”

Section: Introductionmentioning

confidence: 99%

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Cryopreservation: An Overview of Principles and Cell-Specific Considerations

Whaley

Damyar

Witek³

et al. 2021

Cell Transplant

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174

103

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The origins of low-temperature tissue storage research date back to the late 1800s. Over half a century later, osmotic stress was revealed to be a main contributor to cell death during cryopreservation. Consequently, the addition of cryoprotective agents (CPAs) such as dimethyl sulfoxide (DMSO), glycerol (GLY), ethylene glycol (EG), or propylene glycol (PG), although toxic to cells at high concentrations, was identified as a necessary step to protect against rampant cell death during cryopreservation. In addition to osmotic stress, cooling and thawing rates were also shown to have significant influence on cell survival during low temperature storage. In general, successful low-temperature cell preservation consists of the addition of a CPA (commonly 10% DMSO), alone or in combination with additional permeating or non-permeating agents, cooling rates of approximately 1ºC/min, and storage in either liquid or vapor phase nitrogen. In addition to general considerations, cell-specific recommendations for hepatocytes, pancreatic islets, sperm, oocytes, and stem cells should be observed to maximize yields. For example, rapid cooling is associated with better cryopreservation outcomes for oocytes, pancreatic islets, and embryonic stem cells while slow cooling is recommended for cryopreservation of hepatocytes, hematopoietic stem cells, and mesenchymal stem cells. Yields can be further maximized by implementing additional pre-cryo steps such as: pre-incubation with glucose and anti-oxidants, alginate encapsulation, and selecting cells within an optimal age range and functional ability. Finally, viability and functional assays are critical steps in determining the quality of the cells post-thaw and improving the efficiency of the current cryopreservation methods.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Cryopreservation: An Overview of Principles and Cell-Specific Considerations

Whaley

Damyar

Witek³

et al. 2021

Cell Transplant

Self Cite

174

103

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“…The cryopreservation of stem cells and embryos has been successful (10,11). The cryopreservation of rat testicular tissue (12), rat, pig and human islet tissue (13)(14)(15)(16) and liver tissues of mice, rats, dogs, monkeys and humans has also been successful (17). Cryopreservation is not only conducive to the accumulation of donor tissue, but also significantly reduces its immunity (18).…”

Section: Discussionmentioning

confidence: 99%

Comparison of the effects of three cryoprotectants on the cryopreservation of mouse subcutaneous tissue under different conditions

et al. 2020

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The subcutaneous tissue of animals contains different cell types, and different cells have different requirements for cryopreservation. This establishes obstacles that need to be overcome in the clinical application of tissue preservation. In the present study, the effects of different freezing rates and various concentrations of cryoprotectants on the cryopreservation of subcutaneous tissue of mice were compared, and these results provided basic research data that can be used to explore the optimal cryopreservation method for tissue. The effects of three cryoprotectants, dimethyl sulfoxide, glycerinum and 1,2-propanediol, and their concentrations on the cryopreservation of subcutaneous tissue of mice were compared with slow and rapid freezing rates. The results revealed that under various cryopreservation conditions, the percentage of fibroblasts that grow from the tissue following slow cryopreservation (19.8%) was significantly higher than that following rapid freezing (6.7%) at osmotic equilibrium for 10-20 min (P<0.05). After 19 days of culture, under the conditions of slow freezing, with 10, 20 and 30% glycerinum as a cryoprotectant, respectively, fibroblasts grew from 26.0, 16.7 and 16.7% of the tissues, respectively. No fibroblasts were indicated in the tissue mass cultured in any other tissue blocks treated with cryopreservation solutions. Under the condition of rapid freezing, fibroblasts grew from 6.7 and 6.7% tissue blocks of 20% DMSO and 10% glycerinum, respectively, following 19 days of culture. No fibroblasts were identified in the tissue mass cultured in the other tissue blocks treated with cryopreservation solutions, and no fibroblasts were identified in the tissue blocks without osmotic balance before freezing.

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“…Islets treated with permeating CPA mixtures achieved significantly higher yield and viability compared to islets treated with DMSO only. When transplanted into STZ-induced mice, islets treated with DMSO/EG mixtures caused normoglycemia 12 days faster on average than DMSO only-treated islets [68].…”

Section: Permeating Cpa Mixturesmentioning

confidence: 93%

Current Advancements in Pancreatic Islet Cryopreservation Techniques

Rodriguez¹,

Whaley²,

Alexander³

et al. 2020

Cryopreservation - Current Advances and Evaluations

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There have been significant advancements in the research of pancreatic islet transplantations over the past 50 years as a treatment for Type 1 Diabetes Mellitus (T1DM). This work has resulted in hundreds of clinical islet transplantation procedures internationally. One limitation of the procedure includes effective storage techniques during donor-recipient cross-matching following islet isolation from deceased donor. Cryopreservation, which is heavily used in embryology research, has been proposed as a prospective method for pancreatic islet banking to bridge the temporal intervals between donor-recipient matching. The cryopreservation methods currently involve the freezing of islets to subzero (−80/−196°C) temperatures for storage followed by a thawing and warming period, which can be increasingly harmful to islet viability and insulin secretion capabilities. Recent advances in islet cryopreservation technologies have improved outcomes for islet health and survivability during this process. The aim of this chapter is to characterize aspects of the islet cryopreservation method while reviewing current procedural improvements that have led to better outcomes to islet health.

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Improved cryopreservation yield of pancreatic islets using combination of lower dose permeable cryoprotective agents

Cited by 15 publications

References 21 publications

Cryopreservation: An Overview of Principles and Cell-Specific Considerations

Cryopreservation: An Overview of Principles and Cell-Specific Considerations

Comparison of the effects of three cryoprotectants on the cryopreservation of mouse subcutaneous tissue under different conditions

Current Advancements in Pancreatic Islet Cryopreservation Techniques

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