Figure 1.
Macroscopic and microscopic appearance of liver tissue used for hepatocyte isolation from resected surgical specimen and explanted diseased livers.
Representative macroscopic appearances of liver preparations removed during partial hepatectomy (lobectomy right) due to Klatskin-Tumor (A) and a liver explant due to hepatocellular carcinoma (HCC) (B) already showing marked differences in terms of cirrhotic alterations in the latter. The corresponding cut surfaces and representative histologic sections show homogenous liver parenchyma without any significant pathological finding in the former (C, F) and a mixed micro- and macronodular cirrhosis with mild chronic inflammation in the latter (D, G). Several smooth walled cysts are apparent on the cut surface of an explant of a polycystic liver (E). A corresponding histological section shows areas of fibrosis with cystically dilated bile duct structures with intervening areas of inconspicuous liver parenchyma (H).
Table 1.
Demographics and clinical data of tissue donors for cell isolation.
Table 2.
Results of cell isolation.
Figure 2.
Comparable morphological appearance of cultured hepatocytes isolated from resected surgical specimens and explanted diseased livers.
Phase-contrast microscopy of cultured (day 5) primary human hepatocytes isolated from resected surgical specimens (Rx-group) removed due to metastasis of colorectal cancer (CRC) (A), hepatocellular carcinoma (HCC) (B) and cholangiocellular carcinoma (CCC) (C) as well as from explanted diseased livers (Ex-group) due to HCC (D) and polycystic liver disease (PLD) (E). Hepatocytes of both groups show the typical polygonal shape, are highly prismatic and either mono- or polynucleated. Signs of the formation of bile canaliculi are present. Magnification 100x.
Figure 3.
Comparable courses of AST-leakage, albumin synthesis, urea production and in vitro cell number in hepatocytes isolated from resected surgical specimens and explanted diseased livers.
Diagrams depicting the courses of AST-leakage (A), albumin synthesis (B) and urea production (C) determined from culture supernatants on days 1, 3, 5 and 7 (following daily change of the culture medium) for the resection-group (Rx, white circles) and explant-group (Ex, black squares), respectively. Subsequent analysis of the in vitro cell numbers (D) by CyQuant assay was performed using the appropriate cell pellets. Data is presented as mean ±SEM of n = 5 experiments.
Figure 4.
Transcript expression of albumin and CYP genes are comparable in hepatocytes isolated from resected surgical specimens and explanted diseased livers.
Diagram depicting the mRNA-expression of albumin (A) determined by RT-PCR on days 1, 3, 5 and 7 (Rx: light grey bars, Ex: dark grey bars). Furthermore, the courses of basal expression of cytochromes P450 (CYP) 1A1 (B), 2C8 (C) and 3A4 (D) are shown for the resection-group (Rx, light grey bars) and explant-group (Ex, dark grey bars), respectively. Following induction of the 7-ethoxycoumarin-O-deethylase (ECOD) activity by exposing the cells to 3-methylcholantren, rifampicin and phenobarbital for 72 h, courses of mRNA-expression in relation to the unstimulated control are shown in both groups for CYPs 1A1 (white bars), 2C8 (light grey bars) and 3A4 (dark grey bars) (E). Data is presented as mean ±SEM of n = 5 experiments.
Figure 5.
Phase I and II enzyme reactions are comparable in hepatocytes isolated from resected surgical specimens and explanted diseased livers.
Diagrams depicting the activity of the 7-ethoxycoumarin-O-deethylase (ECOD) (A) in PHH of the resection-group (Rx) and explant-group (Ex) regarding the total 7-hydroxycoumarin as a product (black bars) as well as the non-glucuronidated form (free 7-hydroxycoumarin; white bars). Untreated controls were compared to PHH exposed to 3-methylcholantren, rifampicin and phenobarbital for 72 h ( = drug treated). Furthermore, the uracil-5′-diphosphate-glucuronyltransferase (UDP-GT) activity (B) using 4-methylumbelliferone (4-MU) as a substrate is shown for the Rx- (black bar) and Ex-group (white bar), respectively. Data is presented as mean ±SEM of n = 5 experiments.