High recovery of functional islets stored at low and ultralow temperatures

Bhawna Chandravanshi, Anandh Dhanushkodi, Ramesh Bhonde

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

BACKGROUND: Poor recovery of islets upon cryopreservation is the main hurdle in islet banking. Pancreatic islets have a poor antioxidative defense mechanism, and exposure of islets to low temperature leads to oxidative stress. AIM: We aimed to investigate whether known compounds such as metformin, γ aminobutyric acid (GABA), docosahexanoic acid (DHA), or eicosapentaenoic acid (EPA) alone or in combination are capable of reducing oxidative stress for better islet recovery upon storage at suboptimal temperatures. METHODS: Islets isolated from mouse pancreas were stored at low temperature (4°C) for 15 days and at ultralow temperature (-196°C) for 30 days with or without additives. After revival from cold storage, islets were assessed by using three methods: (1) specificity by dithizone (DTZ), (2) viability by fluorescein diacetate/propidium iodide (FDA/PI) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and (3) functionality by glucose- stimulated insulin secretion (GSIS). The oxidative status of the islets stored at suboptimal temperatures was determined by both intracellular free radical release (fluorometric analysis) and lipid peroxidation (enzymatic determination). RESULTS: Supplementation with additives led to an improvement in islet survival upon storage at suboptimal temperatures, without depletion of insulin secretory activity, which was comparable to that of controls. The additives acted as cryoprotectants and antioxidants as revealed by high recovery of viable islets and reduction in total reactive oxygen species (ROS) and malonidealdehyde (MDA), respectively. CONCLUSIONS: Our results demonstrate for the first time that supplementation with EPA, DHA, and metformin may lead to higher islet recovery from -196°C storage, enabling proper islet banking.

Original languageEnglish
Pages (from-to)267-278
Number of pages12
JournalReview of Diabetic Studies
Volume11
Issue number3-4
DOIs
Publication statusPublished - 11-07-2014

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Temperature
Eicosapentaenoic Acid
Metformin
Oxidative Stress
Dithizone
Insulin
Aminobutyrates
Fluorometry
Acids
Propidium
Cryopreservation
Islets of Langerhans
gamma-Aminobutyric Acid
Lipid Peroxidation
Free Radicals
Pancreas
Reactive Oxygen Species
Antioxidants
Glucose

All Science Journal Classification (ASJC) codes

  • Endocrinology, Diabetes and Metabolism
  • Endocrinology
  • Internal Medicine
  • Medicine(all)

Cite this

Chandravanshi, Bhawna ; Dhanushkodi, Anandh ; Bhonde, Ramesh. / High recovery of functional islets stored at low and ultralow temperatures. In: Review of Diabetic Studies. 2014 ; Vol. 11, No. 3-4. pp. 267-278.
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High recovery of functional islets stored at low and ultralow temperatures. / Chandravanshi, Bhawna; Dhanushkodi, Anandh; Bhonde, Ramesh.

In: Review of Diabetic Studies, Vol. 11, No. 3-4, 11.07.2014, p. 267-278.

Research output: Contribution to journalArticle

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N2 - BACKGROUND: Poor recovery of islets upon cryopreservation is the main hurdle in islet banking. Pancreatic islets have a poor antioxidative defense mechanism, and exposure of islets to low temperature leads to oxidative stress. AIM: We aimed to investigate whether known compounds such as metformin, γ aminobutyric acid (GABA), docosahexanoic acid (DHA), or eicosapentaenoic acid (EPA) alone or in combination are capable of reducing oxidative stress for better islet recovery upon storage at suboptimal temperatures. METHODS: Islets isolated from mouse pancreas were stored at low temperature (4°C) for 15 days and at ultralow temperature (-196°C) for 30 days with or without additives. After revival from cold storage, islets were assessed by using three methods: (1) specificity by dithizone (DTZ), (2) viability by fluorescein diacetate/propidium iodide (FDA/PI) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and (3) functionality by glucose- stimulated insulin secretion (GSIS). The oxidative status of the islets stored at suboptimal temperatures was determined by both intracellular free radical release (fluorometric analysis) and lipid peroxidation (enzymatic determination). RESULTS: Supplementation with additives led to an improvement in islet survival upon storage at suboptimal temperatures, without depletion of insulin secretory activity, which was comparable to that of controls. The additives acted as cryoprotectants and antioxidants as revealed by high recovery of viable islets and reduction in total reactive oxygen species (ROS) and malonidealdehyde (MDA), respectively. CONCLUSIONS: Our results demonstrate for the first time that supplementation with EPA, DHA, and metformin may lead to higher islet recovery from -196°C storage, enabling proper islet banking.

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