Cultured human hepatocytes have broad research and clinical applications. However, culturing these cells is difficult.
There can be rapid loss of the hepatocytic phenotype in primary culture, and a limited replicating capacity of the cultured cells.
In this study, scientists from Australia and the United States describe the establishment of serum-free primary cultures of human fetal hepatocytes.
These cultures retained their hepatocytic morphology and gene expression patterns for several months. They also maintained sufficient proliferative activity to permit subculturing for at least 2 passages.
|Cultures retained their hepatocytic morphology and gene expression patterns for several months.|
The team found that the human fetal hepatocyte cultures contained 2 main cell types, which morphologically resembled large and small hepatocytes. The cells expressed alpha-fetoprotein, cytokeratin 19, albumin, and other hepatic proteins.
The scientists determined that treatment of the cultures with oncostatin M increased cell size, and enhanced cell differentiation and formation of bile canaliculi. This may have been due to an effect on hepatocyte nuclear factor 4.
They found that a month after plating, multiple clusters of very small cells became apparent in the cultures. These cells had very few organelles and the team referred to them as blast-like cells.
Flow cytometric analysis of these cells showed that they express oval cell/stem cell markers such as CD90 (Thy-1), CD34, and OV-6, However, they do not stain with antibodies to 2-microglobulin.
The scientists determined that human fetal hepatocyte cultures maintained for 9 to 12 months produced grossly visible organoids. These contained ductular structures that stained for cytokeratin 18 and 19, and alpha-fetoprotein.
Dr Catherine Lázaro's team concluded, "Human fetal hepatocyte cultures…constitute an excellent tool for a variety of studies with human hepatocytes, including the mechanisms of viral infection".