Söderholm J D, Hedman L, Artursson P, Franzén L, Larsson J, Pantzar N, Permert J, Olaison G
Department of Surgery and Clinical Research Center, University Hospital, Linköping, Sweden.
Acta Physiol Scand. 1998 Jan;162(1):47-56. doi: 10.1046/j.1365-201X.1998.0248f.x.
The Ussing chamber is increasingly being used for in vitro studies of human intestinal mucosa, but little attention has been paid to the viability of specimens over time. Ninety-one mucosal specimens from the ileum in 19 patients operated on for colonic cancer were studied in regard to intestinal barrier function, metabolism, electrophysiology and histology during 360 min of incubation in Ussing chambers. Steady-state permeability to 51Cr-EDTA was maintained for 120 min. Mucosal ATP and lactate levels were stable for 180 min and transmucosal glucose flux for 240 min. Lactate dehydrogenase leakage was limited within 120 min. Transepithelial potential difference was 9.0 +/- 3.0 mV at the start, and declined slowly throughout 360 min. Light microscopy revealed epithelial lifting from the basal lamina at 90 min. Transmission electron microscopy demonstrated preserved ultrastructure for 120 min. Specimens with a transepithelial potential difference below 6 mV at the start were associated with increased 51Cr-EDTA permeability and lactate dehydrogenase leakage and more pronounced light microscopy changes. All studied parameters pointed to preserved viability if experiments were kept within a period of 90 min after equilibration. The few specimens with early viability derangement were identified by a transepithelial potential difference below 6 mV at the start. The Ussing chamber provides a tool for in vitro studies of human intestinal epithelium, including permeability. To minimize viability problems, experiments should be limited in time and monitored by measurements of transepithelial potential difference.
尤斯灌流小室越来越多地用于人体肠黏膜的体外研究，但随着时间推移，对标本活力的关注却很少。对19例因结肠癌接受手术的患者的91份回肠黏膜标本，在尤斯灌流小室中孵育360分钟期间，就肠屏障功能、代谢、电生理学和组织学进行了研究。对51Cr-乙二胺四乙酸（51Cr-EDTA）的稳态通透性维持了120分钟。黏膜三磷酸腺苷（ATP）和乳酸水平在180分钟内保持稳定，跨黏膜葡萄糖通量在240分钟内保持稳定。乳酸脱氢酶泄漏在120分钟内受到限制。跨上皮电位差在开始时为9.0±3.0毫伏，并在整个360分钟内缓慢下降。光学显微镜显示在90分钟时上皮从基底层抬起。透射电子显微镜显示超微结构在120分钟内保持完好。开始时跨上皮电位差低于6毫伏的标本，其51Cr-EDTA通透性增加、乳酸脱氢酶泄漏增加，且光学显微镜下变化更明显。如果实验在平衡后90分钟内进行，所有研究参数均表明标本活力得以保留。少数早期活力紊乱的标本在开始时通过跨上皮电位差低于6毫伏得以识别。尤斯灌流小室为人体肠上皮的体外研究，包括通透性研究，提供了一种工具。为尽量减少活力问题，实验应在时间上加以限制，并通过测量跨上皮电位差进行监测。
