MATERIALS AND METHODS INJURY in many s is characterized by , formation of surface blebs, and swelling (22, 24, 34). Disruption of volume regulation may promote bleb formation and the progression to . In ischemic kidney and heart, reperfusion with hyperosmolar mannitol reduced lethal injury, an effect attributed to reversal of swelling and improvement of vascular perfusion (11, 26). anoxia in cultured renal proximal tubular s, polyethylene glycol (PEG), a high-molecular-weight solute, prevented blebbing and improved viability after reoxygenation (19). In cultured hepatocytes, bleb growth accelerated and volume, judged by cross-sectional area, increased rapidly just before the onset of from anoxia and chemical hypoxia (16, 23). Loss of viability was preceded by a lysis of a surface bleb. These studies suggested therefore that swelling may play a critical role in bleb formation and lethal injury. However, HYPOXIC 0363-6143/89 $1.50 Copyright Hepatocyte isolation. Hepatocytes were isolated from 16- to 24-h fasted male Sprague-Dawley rats (200-250 g) by collagenase perfusion as previously described (13, 14, 16). Viability of hepatocytes averaged 94% (range of 88 99%) by trypan blue exclusion. Solutions. The basic incubation media was Krebs Ringer-N-2-hydroxyethylpiperazine-Nâ-2-ethanesulfonic acid (HEPES) (KRH) buffer containing (in mM) 115 NaCl, 5 KCl, 1 KH2P04,
AJP - Cell Physiology – The American Physiological Society
Published: Aug 1, 1989
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