Repair pathways evident in human liver organ slices

Abstract

The extension of human liver slice culture viability for several days broadens the potential of this ex vivo model for characterizing pathways of organ injury and repair, and allows for the multiple dosing of compounds. Extended viability is demonstrated by continued synthesis of GSH and ATP, and maintenance of intracellular K(+) levels. Gene expression profiling revealed the activation of regeneration pathways via increased expression of collagens (I, IV, and V), laminins, ninjurin, growth factors (EGF, epiregulin, and TGF-β1), matrix metalloproteinase-7, and insulin like growth factor 5. Collagen IV protein levels began to increase by day 4 of culture. Some markers of hepatic stellate cells, detected by RT-PCR, were up-regulated (HSP47, αSMA, pro-collagen 1a1, PDGF receptor, thrombospondin-2) with time in culture, while other markers exhibited no change or were down-regulated (αB-crystallin, synaptophysin), suggesting that the induction of regenerative pathways may in part be the role of the stellate cells as well as resident fibroblasts. Complimentary to the gene expression was evidence of regeneration in the human liver slices, as evaluated by histopathology. Improvements in organ acquisition, organ slice preparation and culture methods demonstrates that organ slice viability, integrity and morphology can be extended reproducibly for several days in culture which allows for the investigation of injury and repair processes.