In the present study, the developmental expression of three cytochrome P450 (P450) isoforms, 1A1, 2B1 and 3A2, and the ability to store glycogen was investigated in intrasplenic liver cell explants in comparison to adult and fetal liver. Fetal liver tissue suspensions were transplanted into the spleens of adult male syngenic Fisher inbred rats. Animals were sacrificed at 3 days, 1, 2, 4 weeks, 2, 4, 6 months and 1 year after transplantation. Spleens and livers of transplant recipients were compared to those of sham operated and control rats. Three days after transplantation little bulks of hepatocytes and only few bile ducts were seen in the red pulp of the transplant containing spleens. A massive hypertrophy and proliferation of bile ducts and also an augmentation in the number of hepatocytes were observed 4 weeks after transplantation. One month later, however, the bile ducts had become more and more atrophic, while instead the number of hepatocytes continuously increased. One year after surgery large masses of hepatocytes with apparent cord structure and only few but well preserved bile ducts were seen. Within the livers of adult rats, P450 1A1 was only slightly expressed by some hepatocytes around the central veins. P450 2B1 and 3A2 isoforms expression was much stronger, but also predominantly located in the hepatocytes of the central zone of the liver lobule. Hepatocytes of fetal livers displayed a moderate P450 1A1 expression. In some cells also a very mild staining for P450 2B1 and 3A2 was observed. Within the hepatocytes of the intrasplenic liver cell explants P450 1A1 was still expressed 3 days after transplantation, disappeared at 1 week after surgery, but reappeared at 4 weeks after transplantation. After 2, 4 and 6 months no staining for P450 1A1 was detectable any more. One year after transplantation again a slight P450 1A1 expression appeared. With P450 2B1 and 3A2 a mild to moderate expression was seen already at 3 days after transplantation. Four weeks after surgery nearly all of the hepatocytes were stained for P450 2B1 and 3A2, but there were marked differences between the individual cells in the extent of the expression of these two P450 subtypes, like it was also the case with normal adult liver. Within hepatocytes of the fetal livers strongly stained glycogen granules were seen, which, in comparison to adult livers, were rather coarse-grained. Three days after transplantation the glycogen granules in the transplanted hepatocytes were still coarse-grained, but from 1 week after transplantation on, they became more and more fine-grained. As it was also the case with normal adult liver cells, there were marked differences between the individual transplanted hepatocytes in their glycogen content. These results demonstrate that transplanted liver cells originating from syngenic fetal liver tissue suspensions can survive in host organs like the spleen for at least 1 year. They proliferate, differentiate, are able to store glycogen, and express different P450 isoforms, like normal adult liver cells.