Are preoperative laboratory parameters predictive for the metabolic function of primary human hepatocytes?

Background:

Primary human hepatocytes are used for a variety of experiments in basic and clinical research. There is a substantial amount of literature available on which liver specimen is the most promising for a high cell yield and viability. New approaches such as the „organ-on-a-chip“ or the de- and recellularization of organs require hepatocytes of high quality as well as good cell yield. We investigated if patient's clinical parameters could be used as markers to predetermine viability and in-vitro metabolic activity of isolated human hepatocytes. Thus, we focused on identifying markers which might impair the comparability of experimental groups where hepatocytes were used from heterogenous hepatocyte sources.

Methods:

Primary human hepatocytes were isolated using a two-step collagenase perfusion protocol. Hepatocytes were then cultured for seven days in six-well plates. Clinical chemistry samples were taken and analyzed on day 1, 2, 4 and 6. Cytochrome P450 enzymes CYP 1A2, CYP 2B6, CYP 3A4 were analyzed. Patient data was extracted from preoperative case files and routine laboratory testing of 39 patients.

Results:

We found that high levels of preoperative bilirubin and Gamma-glutamyltransferase – mostly found in cholestatic livers – had a significant impact on the metabolic function of the isolated hepatocytes. There has been a strong correlation between preoperative parameters of bilirubin and Gamma-glutamyltransferase and in-vitro parameters of AST, urea and albumine. Other parameters taken preoperatively did not significantly affect the metabolic function of isolated hepatocytes.

Discussion:

Our results show that preoperative laboratory parameters can be used to predict the metabolic function of primary human hepatocytes in culture. These parameters can be used to decide which specimen should be used for isolation.