An organ culture of postnatal rat liver slices

Abstract

A technique for the organ culture of postnatal and adult rat liver has been developed. Liver slices, 0.3 mm thick, were maintained in Conway units at the interphase between medium and a 95% O2:5% CO2 atmosphere. Postnatal liver in culture for up to 72 h had healthy hepatocytes throughout the explants; if adult liver was used the upper 0.2 mm was healthy after 24 h. These slices incorporated tritiated orotate and leucine into trichloroacetic acid-precipitable material. Incorporation of orotate was shown to be spread over the entire slice of neonatal liver. Culturing did not alter the potassium ion content of postnatal liver. Tyrosine aminotransferase activity in liver slices from postnatal, adult, and adrenalectomized adult rats was stimulated by glucocorticoids and dibutyryl cyclic AMP. Cycloheximide and actinomycin D prevented this response. Further, cortisol exerted a permissive effect on the stimulation of tyrosine aminotransferase activity by dibutyryl cyclic AMP in slices from adrenalectomized rats. Induction of urea cycle enzymes by cortisol was demonstrated in cultures of liver from adrenalectomized adult animals.

Long-Term Organotypic Slice Culture of the Neonatal Mouse Liver.

“…Three culture systems have been reported to study hepatocytes or cholangium cells in vitro; primary monolayer [1,11], spheroid [10,13], and organ cultures [9] . Monolayer cultures of isolated parenchymal cells are frequently used for in vitro study [2].…”

Hormone‐ and endotoxin‐modulated gene expression of a long‐term organ culture system of adult rat liver

“…In some cases a pretreatment of the animals has yielded 'liver-specific' in vitro results (e.g, with LPS [15], aroclor 1254 [16], phenobarbital [17] or adrenalectomy [18]). These approaches do not provide an alternative for studies requiring large numbers of laboratory animals.…”

Hormonal Regulation of Four Urea Cycle Enzymes in Postnatal Rat Liver in Organ Culture

“…Then the serum component was added. The viability of liver slices prepared by this method has been established by Hart et al [14], Slices in culture for up to 72 h had healthy hepatocytes throughout, incorporated tritia ted orotate and had unaltered intracellular potassium ion content [14], Liver homogenates in 20 mmol/1 potassium phos phate buffer, pH 7.4, and dilutions in 10 mmol/1 man ganese sulfate were prepared for the urea cycle enzyme assays as described previously [2]. CPS, OTC, ASS,…”

“…Thus, we used an organ culture of 13-day-old rat liver, devel oped by us [14], to explore further the hor monal factors which may trigger the increases in the urea cycle enzyme levels during the 3rd week of life and to establish the direct in fluence of hormones on this tissue at this cru cial time in postnatal development. Four urea cycle enzymes were studied: CPS, OTC, ASS, and arginase.…”

“…The organ culture technique was that described by Hart et al [14], Culture medium consisted of 50% synthetic component (medium 199 containing 0.35 g/1 sodium bicarbonate, 50 mg/1 streptomycin sulfate and 5 X IO4 U/l penicillin) and 50% fetal bovine serum. Cultures were prepared using the livers of 6-8 13-dayold litter-mates of similar weights.…”