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KMID : 1147720100030040241
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Journal of Acupuncture and Meridian Studies
2010 Volume.3 No. 4 p.241 ~ p.248
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Viability Assessment of Primo-node Slices From Organ Surface Primo-vascular Tissues in Rats
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Han Tae-Hee
Lim Chae-Jeong
Choi Jae-Hong
Lee So-Yeong
Ryu Pan-Dong
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Abstract
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The primo-vascular system is a novel thread-like structure which is recently rediscovered, but its cellular properties are largely unknown. In this study, a slice preparation for primo-nodes was developed to facilitate study of the cellular properties of primo-node cells in vitro. Slices (4?8 slices; 200 μm thick) were sectioned from single primo-nodes collected from the abdominal organ surface of rats and incubated in oxygenated Krebs solution at 25°C or 31°C for up to 7 hours. Trypan blue staining and whole-cell patch-clamp recordings were performed to estimate the viability of cells in the slices. Viability was largely maintained during the first 3 hours, but subsequently decreased (from 80% to 21%, p < 0.001). In addition, the viability of slices incubated at 31°C was higher than those incubated at 25°C (80% vs. 47%, p < 0.001). In whole-cell patch-clamp experiments, high resistance seals readily formed and primo-node cells showed a mean resting membrane potential (-38 mV) comparable to that recorded with sharp electrodes and outwardly-rectifying current-voltage relationships. The results show that the primo-node slices developed in this study maintained viability for up to 4 hours in vitro.
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KEYWORD
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Bonghan corpuscle, colorimetric assays, novel circulatory system, slice patch clamp
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