title: Viability assessment of primo-node slices from organ surface primo-vascular tissues in rats.
abstract: 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.
authors.affiliation: Laboratory of Veterinary Pharmacology, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul, Korea.
authors.forename: Tae Hee
authors.initials: TH
authors.lastname: Han
authors.forename: Chae Jeong
authors.initials: CJ
authors.lastname: Lim
authors.forename: Jae-Hong
authors.initials: JH
authors.lastname: Choi
authors.forename: So Yeong
authors.initials: SY
authors.lastname: Lee
authors.forename: Pan Dong
authors.initials: PD
authors.lastname: Ryu
mesh_terms: D015669:Acupuncture Points; D000825:Animal Structures / Q000033:anatomy & histology / Q000737:chemistry / Q000502:physiology*; D000818:Animals; D016591:Histocytological Preparation Techniques / Q000379:methods*; D008297:Male; D008564:Membrane Potentials; D008867:Microtomy; D051381:Rats; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling