Luminescence technology in preservation and transplantation for rat islet

Negishi, K; Teratani, Takumi; Iwasaki, Junji; Kanazawa, Hideko; Kasahara, Naoya; Lefor, Alan Kawarai; Üemoto, Shinji; Fujimoto, Yasuhiro; Kobayashi, Eiji

doi:10.4161/isl.3.3.15626

Cited by 14 publications

(12 citation statements)

References 25 publications

(30 reference statements)

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“…Therefore, this system shows good performance for screening the effect of different factors in a preservation medium by measuring their results on the luciferase activity as an index of the remaining amount of ATP. To date, this method has been used to screen several types of preservation media in various organs such as the heart16, liver17, kidney18, islets1920, and small intestine21. However, to our knowledge, a preservation medium that is ideal for tissue-engineered cell sheets has not yet been screened.…”

mentioning

confidence: 99%

Ebselen Preserves Tissue-Engineered Cell Sheets and their Stem Cells in Hypothermic Conditions

Katori

Hayashi

Kobayashi

et al. 2016

Sci Rep

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Clinical trials have been performed using autologous tissue-engineered epithelial cell sheets for corneal regenerative medicine. To improve stem cell-based therapy for convenient clinical practice, new techniques are required for preserving reconstructed tissues and their stem/progenitor cells until they are ready for use. In the present study, we screened potential preservative agents and developed a novel medium for preserving the cell sheets and their stem/progenitor cells; the effects were evaluated with a luciferase-based viability assay. Nrf2 activators, specifically ebselen, could maintain high ATP levels during preservation. Ebselen also showed a strong influence on maintenance of the viability, morphology, and stem cell function of the cell sheets preserved under hypothermia by protecting them from reactive oxygen species-induced damage. Furthermore, ebselen drastically improved the preservation performance of human cornea tissues and their stem cells. Therefore, ebselen shows good potential as a useful preservation agent in regenerative medicine as well as in cornea transplantation.

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confidence: 99%

Ebselen Preserves Tissue-Engineered Cell Sheets and their Stem Cells in Hypothermic Conditions

Katori

Hayashi

Kobayashi

et al. 2016

Sci Rep

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“…Since light emission of luciferase-positive cells requires ATP and oxygen, the luciferase-luciferin reaction can be used to determine the metabolic activity and ATP levels in the cells (13,29). The photon intensity of cold-preserved islets has been shown to correlate with islet viability (29). In this study, the intensity of bioluminescence decreased during the 6 h of rewarming after 18 h of cold preservation, whereas no change was observed in islets without cold preservation (Fig.…”

Section: Resultsmentioning

confidence: 75%

“…The effect of cold preservation/rewarming on the metabolic activity of isolated islets was examined using Firefly rat islets and bioluminescence imaging. Since light emission of luciferase-positive cells requires ATP and oxygen, the luciferase-luciferin reaction can be used to determine the metabolic activity and ATP levels in the cells (13,29). The photon intensity of cold-preserved islets has been shown to correlate with islet viability (29).…”

Section: Resultsmentioning

confidence: 99%

Involvement of a proapoptotic gene (BBC3) in islet injury mediated by cold preservation and rewarming

Omori

Kobayashi

Komatsu

et al. 2016

American Journal of Physiology-Endocrinology and Metabolism

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Long-term pancreatic cold ischemia contributes to decreased islet number and viability after isolation and culture, leading to poor islet transplantation outcome in patients with type 1 diabetes. In this study, we examined mechanisms of pancreatic cold preservation and rewarming-induced injury by interrogating the proapoptotic gene BBC3/Bbc3, also known as Puma (p53 upregulated modulator of apoptosis), using three experimental models: 1) bioluminescence imaging of isolated luciferase-transgenic (“Firefly”) Lewis rat islets, 2) cold preservation of en bloc-harvested pancreata from Bbc3-knockout (KO) mice, and 3) cold preservation and rewarming of human pancreata and isolated islets. Cold preservation-mediated islet injury occurred during rewarming in “Firefly” islets. Silencing Bbc3 by transfecting Bbc3 siRNA into islets in vitro prior to cold preservation improved postpreservation mitochondrial viability. Cold preservation resulted in decreased postisolation islet yield in both wild-type and Bbc3 KO pancreata. However, after culture, the islet viability was significantly higher in Bbc3-KO islets, suggesting that different mechanisms are involved in islet damage/loss during isolation and culture. Furthermore, Bbc3-KO islets from cold-preserved pancreata showed reduced HMGB1 (high-mobility group box 1 protein) expression and decreased levels of 4-hydroxynonenal (4-HNE) protein adducts, which was indicative of reduced oxidative stress. During human islet isolation, BBC3 protein was upregulated in digested tissue from cold-preserved pancreata. Hypoxia in cold preservation increased BBC3 mRNA and protein in isolated human islets after rewarming in culture and reduced islet viability. These results demonstrated the involvement of BBC3/Bbc3 in cold preservation/rewarming-mediated islet injury, possibly through modulating HMGB1- and oxidative stress-mediated injury to islets.

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“…Fluorescence intensity of kidney samples after preservation was determined at specified time points using a plate reader (Mithras LB940, Berthold Japan, Tokyo, Japan) as previously described [46]. The plate reader was placed in the cold room, and the fluorescence intensity was measured at 4°C.…”

Section: Methodsmentioning

confidence: 99%

Impact of Normothermic Preservation with Extracellular Type Solution Containing Trehalose on Rat Kidney Grafting from a Cardiac Death Donor

et al. 2012

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Background The aim of this study was to investigate factors that may improve the condition of a marginal kidney preserved with a normothermic solution following cardiac death (CD) in a model of rat kidney transplantation (RTx). Methods Post-euthanasia, Lewis (LEW) donor rats were left for 1 h in a 23°C room. These critical kidney grafts were preserved in University of Wisconsin (UW), lactate Ringer's (LR), or extracellular-trehalose-Kyoto (ETK) solution, followed by intracellular-trehalose-Kyoto (ITK) solution at 4, 23, or 37°C for another 1 h, and finally transplanted into bilaterally nephrectomized LEW recipient rats (n = 4–6). Grafts of rats surviving to day 14 after RTx were evaluated by histopathological examination. The energy activity of these marginal rat kidneys was measured by high-performance liquid chromatography (HPLC; n = 4 per group) and fluorescence intensity assay (n = 6 per group) after preservation with UW or ETK solutions at each temperature. Finally, the transplanted kidney was assessed by an in vivo luciferase imaging system (n = 2). Results Using the 1-h normothermic preservation of post-CD kidneys, five out of six recipients in the ETK group survived until 14 days, in contrast to zero out of six in the UW group ( p <0.01). Preservation with ITK rather than ETK at 23°C tended to have an inferior effect on recipient survival ( p = 0.12). Energy activities of the fresh donor kidneys decreased in a temperature-dependent manner, while those of post-CD kidneys remained at the lower level. ETK was superior to UW in protecting against edema of the post-CD kidneys at the higher temperature. Luminescence intensity of successful grafts recovered within 1 h, while the intensity of grafts of deceased recipients did not change at 1 h post-reperfusion. Conclusions Normothermic storage with extracellular-type solution containing trehalose might prevent reperfusion injury due to temperature-dependent tissue edema.

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Luminescence technology in preservation and transplantation for rat islet

Cited by 14 publications

References 25 publications

Ebselen Preserves Tissue-Engineered Cell Sheets and their Stem Cells in Hypothermic Conditions

Ebselen Preserves Tissue-Engineered Cell Sheets and their Stem Cells in Hypothermic Conditions

Involvement of a proapoptotic gene (BBC3) in islet injury mediated by cold preservation and rewarming

Impact of Normothermic Preservation with Extracellular Type Solution Containing Trehalose on Rat Kidney Grafting from a Cardiac Death Donor

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