Evaluation of energy state of islet independent of size using a newly developed ATP bioluminescence assay.
ATP content or energy charge (EC) of islets may be a good parameter to assess viability. In this study, we examined adenine nucleotides and EC of freshly isolated or 24-hour cultured rat islets of various diameters using a novel bioluminescent enzymatic cycling assay system. For freshly isolated islets, ATP content and islet diameter showed a high correlation (r = 0.842, P < .001), but a significant correlation was not observed for cultured islets (r = 0.284) when all islets were included for the analysis. When only the cultured islets with a diameter <350 microm were included for analysis, a significant correlation was observed (r = 0.719). EC of freshly isolated islets fluctuated widely irrespective of diameter, in contrast with results of 24-hour cultured islets, which showed a stable, high EC, regardless of diameter. These data suggest that the ATP content of islets correlates with the islet size and that EC of islets widely fluctuates following isolation, indicating a significant role of monitoring ATP and EC of islets before transplantation.
Ishii
S
S
Department of Surgery I, Fukushima Medical University, 1 Hikarigaoka, Fukushima 960-1295, Japan.
Saito
T
T
Ise
K
K
Sato
Y
Y
Tsutiya
T
T
Kenjo
A
A
Kimura
T
T
Anazawa
T
T
Suzuki
S
S
Terashima
M
M
Gotoh
M
M
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D018929:Cell Culture Techniques; D048429:Cell Size; D004734:Energy Metabolism / Q000502:physiology*; D007515:Islets of Langerhans / Q000166:cytology / Q000502:physiology*; D008163:Luminescent Measurements; D008297:Male; D051381:Rats; D017208:Rats, Wistar
D000255:Adenosine Triphosphate