Establishment, characterization, and long-term maintenance of cultures of human fetal hepatocytes.

摘要: Cultured human hepatocytes have broad research and clinical applications; however, the difficulties in culturing rodent are well known. These problems include rapid loss of hepatocytic phenotype primary culture limited replicating capacity cultured cells. We describe establishment serum-free cultures fetal (HFHs) that retain morphology gene expression patterns for several months maintain sufficient proliferative activity to permit subculturing at least 2 passages. Initially, HFH contained main cell types morphologically resembled large small hepatocytes. The expressed α-fetoprotein (AFP), cytokeratin (CK) 19, albumin, other hepatic proteins. Treatment with oncostatin M (OSM) increased size enhanced differentiation formation bile canaliculi, probably through an effect on hepatocyte nuclear factor (HNF) 4α. Approximately 1 month after plating, multiple clusters very cells became apparent cultures. had few organelles referred as blast-like Flow cytometric analysis these showed they express oval cell/stem markers such CD90 (Thy-1), CD34, OV-6 but do not stain antibodies β2-microglobulin. maintained 9 12 produced grossly visible organoids containing ductular structures stained CK18, CK19, AFP. In conclusion, cultures, which might contain a population stem cells, constitute excellent tool variety studies hepatocytes, including mechanisms viral infection.