[Preservation of hepatocytes of the rat by active transport of glycine].
The measurement of the active accumulation of specific substrates against a concentration gradient in tissue fragments in vitro has proved to be a simple but excellent procedure to ascertain the viability of several cell types and, in parallel with routine microscopy, has become a reliable tool in assaying the biochemical and functional modifications of a tissue under physiopathological conditions. It also has the added advantage of using tissues that are not pretreated in any way such as the preparative procedures used in cell isolation methods. The majority of investigations employing hepatocytes has been carried out in material derived from cell cultures or on isolated cells in suspension. The aim of this study was to determine if active transport could be determined in whole-tissue fragments of livers. Following identification of the specific substrate system and ideal incubation conditions, this method was then used to test the validity of different organ preservation solutions after cold storage in vitro. Livers were rapidly removed from rats and incubated for 1 hour at 37 degrees C in oxygenated Krebs bicarbonate buffer containing the substrates in question marked with a tracer dose of 14C. Preliminary investigations showed that active transport by rat hepatocytes was non-existent for beta-methyl-glucoside, phenylalanine and AIB; low for arginine but very important for glycine and serine and that this uptake could be significantly abolished by the presence of choline in the incubation medium.(ABSTRACT TRUNCATED AT 250 WORDS)
Winistörfer
B
B
Division de chirurgie expérimentale, CHUV, Lausanne.
Mirkovitch
V
V
D000818:Animals; D001693:Biological Transport, Active / Q000502:physiology; D002470:Cell Survival / Q000502:physiology*; D015925:Cryopreservation / Q000379:methods; D005998:Glycine / Q000378:metabolism*; D008099:Liver / Q000166:cytology*; D016031:Liver Transplantation / Q000502:physiology*; D009926:Organ Preservation / Q000379:methods*; D051381:Rats; D013997:Time Factors
D005998:Glycine