BILE ACID CONJUGATION IN HUMAN FETAL LIVER IN ORGAN 207 CULTURE

Abstract

An organ culture system for prolonged maintenance of human fetal liver in vitro has been developed. Using this system, bile acid metabolism was investigated. Multiple liver specimens were obtained from human abortuses and stillbirths ranging from 9-30 wks. gestation(gest.). Within 3hrs. of hysterotomy, 1-2mm pieces of liver were placed in scored petri dishes and incubated in modified Leibovitz medium under air on a rocker panel at 37°C. Morphological integrity was demonstrated by LM and EM for periods up to 10 days in vitro. Functional viability was established by adding 14C-cholic acid to the medium and assaying the conjugates formed after extraction and TLC. Cholic acid(C)was taken up by the tissues and conjugated to form taurocholate(TC)and glycocholate(GC) which were then secreted into the medium at a constant rate during 10 days in vitro and in constant increments during a selected 24hr. period. TC is the predominant conjugate formed by fetuses < 20wks. gest. and with increasing gest. age the proportion of GC synthesized increases. Taurine added to the medium in concentrations equimolar to glycine enhanced TC synthesis at all gest. ages (p<0.001), increasing total conjugate synthesis. The addition of 2×10⁻⁴mM hydrocortisone(HC)to the medium increased the proportion of GC synthesized during early gest.(p=.01). The results establish that in the human fetus taurine is preferentially conjugated with primary bile acid and suggest that HC has a maturation effect on human fetal hepatocytes in organ culture.

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