Oxygenated thawing and rewarming alleviate rewarming injury of cryopreserved pancreatic islets

Komatsu, Hirotake; Barriga, Alyssa; Medrano, Leonard; Omori, Keiko; Kandeel, Fouad; Mullen, Yoko

doi:10.1016/j.bbrc.2017.03.134

Cited by 8 publications

(9 citation statements)

References 23 publications

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“…A hundred islets were obtained for RNA isolation before (precultured) and after (postcultured) 3-day culture in the 2 different O 2 environments, followed by real-time polymerase chain reaction, as described previously. 32 Relative quantities of each transcript were normalized to an endogenous housekeeping gene (ACTB) and expressed as fold increase to precultured. All primers for SLC2A1 (Rn01417099_…”

Section: Gene Expression Analysismentioning

confidence: 99%

Posttransplant oxygen inhalation improves the outcome of subcutaneous islet transplantation: A promising clinical alternative to the conventional intrahepatic site

Komatsu

Rawson

Barriga

et al. 2018

American Journal of Transplantation

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Subcutaneous tissue is a promising site for islet transplantation, due to its large area and accessibility, which allows minimally invasive procedures for transplantation, graft monitoring, and removal of malignancies as needed. However, relative to the conventional intrahepatic transplantation site, the subcutaneous site requires a large number of islets to achieve engraftment success and diabetes reversal, due to hypoxia and low vascularity. We report that the efficiency of subcutaneous islet transplantation in a Lewis rat model is significantly improved by treating recipients with inhaled 50% oxygen, in conjunction with prevascularization of the graft bed by agarose-basic fibroblast growth factor. Administration of 50% oxygen increased oxygen tension in the subcutaneous site to 140 mm Hg, compared to 45 mm Hg under ambient air. In vitro, islets cultured under 140 mm Hg oxygen showed reduced central necrosis and increased insulin release, compared to those maintained in 45 mm Hg oxygen. Six hundred syngeneic islets subcutaneously transplanted into the prevascularized graft bed reversed diabetes when combined with postoperative 50% oxygen inhalation for 3 days, a number comparable to that required for intrahepatic transplantation; in the absence of oxygen treatment, diabetes was not reversed. Thus, we show oxygen inhalation to be a simple and promising approach to successfully establishing subcutaneous islet transplantation.

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Section: Gene Expression Analysismentioning

confidence: 99%

Posttransplant oxygen inhalation improves the outcome of subcutaneous islet transplantation: A promising clinical alternative to the conventional intrahepatic site

Komatsu

Rawson

Barriga

et al. 2018

American Journal of Transplantation

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“…Moreover, exogenously implanted islets rapidly develop hypoxia regardless of their anatomical location(66,67), reflecting an inability of neo-vascularisation to counteract islet O consumption. Attempts to address this problem have included the use of permeably culture supports, such as silicone membranes(18,304) perfluorocarbon (PFC)-based culture dishes(166) or O 2 leaching materials(96), as well as culture under hyperoxic ambient PO 2 (50 kPa)(309). Using a PFC-PDMS construct, Fraker and colleagues elegantly demonstrated that the combination of high substrate solubility and lower As such, a detailed understanding of their physiological environment is of clinical relevance.…”

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confidence: 99%

Defining Physiological Normoxia for Improved Translation of Cell Physiology to Animal Models and Humans

Keeley

Mann

2019

Physiological Reviews

244

204

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The extensive oxygen gradient between the air we breathe (Po2 ~21 kPa) and its ultimate distribution within mitochondria (as low as ~0.5–1 kPa) is testament to the efforts expended in limiting its inherent toxicity. It has long been recognized that cell culture undertaken under room air conditions falls short of replicating this protection in vitro. Despite this, difficulty in accurately determining the appropriate O2 levels in which to culture cells, coupled with a lack of the technology to replicate and maintain a physiological O2 environment in vitro, has hindered addressing this issue thus far. In this review, we aim to address the current understanding of tissue Po2 distribution in vivo and summarize the attempts made to replicate these conditions in vitro. The state-of-the-art techniques employed to accurately determine O2 levels, as well as the issues associated with reproducing physiological O2 levels in vitro, are also critically reviewed. We aim to provide the framework for researchers to undertake cell culture under O2 levels relevant to specific tissues and organs. We envisage that this review will facilitate a paradigm shift, enabling translation of findings under physiological conditions in vitro to disease pathology and the design of novel therapeutics.

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“…53 Since ATP could be made aerobically through oxidative phosphorylation, Komatsu et al hypothesized that the ATP depletion during rewarming could be reduced by using increased oxygen in the chamber around the islets during cryopreservation. 54 Human islets Komatsu et al tested this hypothesis using human islets in two groups, a low oxygen group (21%), and high oxygen group (50%). 54 500 IEQ of human islets were cryopreserved using the islet cryopreservation solution (ICS) for a period of at least three months.…”

Section: Oxygen Environmentmentioning

confidence: 99%

“…54 Human islets Komatsu et al tested this hypothesis using human islets in two groups, a low oxygen group (21%), and high oxygen group (50%). 54 500 IEQ of human islets were cryopreserved using the islet cryopreservation solution (ICS) for a period of at least three months. For oxygenated thawing, the islets were equilibrated under 50% O 2 , 45% N 2 , and 5% CO 2 .…”

Section: Oxygen Environmentmentioning

confidence: 99%

“…Once thawed, they were placed in a 24 well plate, at 250 IEQ/well on ice, and transferred to a 50% oxygen incubator. 54 The islets were incubated for a total of 90 minutes, the first 45 of which at 22 C, and the second 45 minutes at 37 C. They then were analyzed for islet volume and compared to their volume prior to the cryopreservation. They defined this as short-term recovery rate.…”

Section: Oxygen Environmentmentioning

confidence: 99%

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Systematic review of islet cryopreservation

Kojayan

Alexander

Imagawa

et al. 2018

Islets

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Pancreatic islet transplantation is being extensively researched as an alternative treatment for type 1 diabetic patients. This treatment is currently limited by temporal mismatch, between the availability of pancreas and isolated islets from deceased organ donor, and the recipient's need for freshly isolated islets. To solve this issue, cryopreservation of islets may offer the potential to bank islets for transplant on demand. Cryopreservation, however, introduces an overwhelmingly harsh environment to the ever-so-fragile islets. After exposure to the freezing and thawing, islets are usually either apoptotic, non-functional, or non-viable. Several studies have proposed various techniques that could lead to increased cell survival and function following a deep freeze. The purpose of this article is to critically review the techniques of islet cryopreservation, with the goal of highlighting optimization parameters that can lead to the most viable and functional islet upon recovery and/or transplant.

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Oxygenated thawing and rewarming alleviate rewarming injury of cryopreserved pancreatic islets

Cited by 8 publications

References 23 publications

Posttransplant oxygen inhalation improves the outcome of subcutaneous islet transplantation: A promising clinical alternative to the conventional intrahepatic site

Posttransplant oxygen inhalation improves the outcome of subcutaneous islet transplantation: A promising clinical alternative to the conventional intrahepatic site

Defining Physiological Normoxia for Improved Translation of Cell Physiology to Animal Models and Humans

Systematic review of islet cryopreservation

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