Isolation and characterization of a polarized isolated hepatocyte preparation in the skateRaja erinacea

✍ Scribed by Smith, D. J. ;Grossbard, M. ;Gordon, E. R. ;Boyer, J. L.

• Publisher: John Wiley and Sons
• Year: 1987
• Tongue: English
• Weight: 606 KB
• Volume: 241
• Category: Article
• ISSN: 0022-104X
• DOI: 10.1002/jez.1402410303

✦ Synopsis

Hepatocytes of the small skate (Raja erinacea) were isolated by collagenase perfusion and evaluated by a variety of functional and morphologic criteria. Cell yield was 1.45 x 10' f 1.3 x lo7 cells per isolation, and as long as 8 h after isolation 98% of the hepatocytes excluded Trypan blue and no leakage of lactate dehydrogenase (LDH) or cell associated potassium could be detected. Oxygen consumption averaged 1.6 k 0.5 nmol/min/mg cell protein, was not stimulated by 1 mM succinate, and also remained stable for up to 8 h following isolation. However, 2,4,-dinitrophenol (5 x lop5 M) produced a 55% increase in oxygen utilization while ouabain, (1 mM) or sodium removal decreased oxygen consumption by 31 f 6 and 33 & 7%, respectively, indicating that a significant portion of the cells energy utilization is coupled to the activity of plasma membrane Naf , Kf -ATPase.