Viability of the Myocardium after Twenty-four-hour Heart Conservation - a Preliminary Study

Improvements in pancreatic islet transplantation for treatment of diabetes are hindered by the absence of meaningful islet quality assessment methods. Oxygen consumption rate (OCR) has previously been used to assess the quality of organs and primary tissue for transplantation. In this study, we describe and characterize a stirred microchamber for measuring OCR with small quantities of islets. The device has a titanium body with a chamber volume of about 200 µL and is magnetically stirred and water jacketed for temperature control. Oxygen partial pressure (pO 2 ) is measured by fluorescence quenching with a fiber optic probe, and OCR is determined from the linear decrease of pO 2 with time. We demonstrate that measurements can be made rapidly and with high precision. Measurements with βTC3 cells and islets show that OCR is directly proportional to the number of viable cells in mixtures of live and dead cells and correlate linearly with membrane integrity measurements made with cells that have been cultured for 24 h under various stressful conditions.

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A stirred microchamber for oxygen consumption rate measurements with pancreatic islets

Papas

Pisania

et al. 2007

Biotech & Bioengineering

102

106

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Islet assessment for transplantation

Papas

Suszynski

Colton

2009

Current Opinion in Organ Transplantation

105

132

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Purpose of review-There is a critical need for meaningful viability and potency assays that characterize islet preparations for release prior to clinical islet cell transplantation (ICT). Development, testing, and validation of such assays have been the subject of intense investigation for the past decade. These efforts are reviewed, highlighting the most recent results while focusing on the most promising assays.Recent Findings-Assays based on membrane integrity do not reflect true viability when applied to either intact islets or dispersed islet cells. Assays requiring disaggregation of intact islets into individual cells for assessment introduce additional problems of cell damage and loss. Assays evaluating mitochondrial function, specifically mitochondrial membrane potential, bioenergetic status, and cellular oxygen consumption rate (OCR), especially when conducted with intact islets, appear most promising in evaluating their quality prior to ICT. Prospective, quantitative assays based on measurements of OCR with intact islets have been developed, validated and their results correlated with transplant outcomes in the diabetic nude mouse bioassay.Conclusion-More sensitive and reliable islet viability and potency tests have been recently developed and tested. Those evaluating mitochondrial function are most promising, correlate with transplant outcomes in mice, and are currently being evaluated in the clinical setting.

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Viability of the Myocardium after Twenty-four-hour Heart Conservation - a Preliminary Study

Abstract: The data indicate full viability of the myocardium after 24 h conservation. The preservation technique described could contribute to the extension of conservation times in heart transplantation and enable transplantation of marginal donor hearts.

Cited by 2 publications

References 12 publications

A stirred microchamber for oxygen consumption rate measurements with pancreatic islets

A stirred microchamber for oxygen consumption rate measurements with pancreatic islets

Islet assessment for transplantation

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