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Hepatic progenitor cells (HPCs) are a potential cell source for liver cell transplantation but do not function like mature liver cells. We sought an effective and reliable method to induce HPC maturation. An immortalized HP14.5 albumin promoter-driven Gaussian luciferase (ALB-GLuc) cell line was established from HPCs isolated from fetal mouse liver of post coitus day 14.5 mice to investigate the effect of induction factors on ALB promoter. HP14.5 parental cells were cultured in DMEM with different combinations of 2% horse serum (HS), 0.1 µM dexamethasone (DEX), 10 ng/mL hepatic growth factor (HGF), and/or 20 ng/mL fibroblast growth factor 4 (FGF4). Trypan blue and crystal violet staining were used to assess cell proliferation with different induction conditions. Expression of hepatic markers was measured by semi-quantitative RT-PCR, Western blot, and immunofluorescence. Glycogen storage and metabolism were detected by periodic acid-Schiff and indocyanine green (ICG) staining. GLuc activity indicated ALB expression. The combination of 2% HS+0.1 µM Dex+10 ng/mL HGF+20 ng/mL FGF4 induced the highest ALB-GLuc activity. Cell proliferation decreased in 2% HS but increased by adding FGF4. Upon induction, and consistent with hepatocyte development, DLK, AFP, and CK19 expression decreased, while ALB, CK18, and UGT1A expression increased. The maturity markers tyrosine aminotransferase and apolipoprotein B were detected at days 3 and 6 post-induction, respectively. ICG uptake and glycogen synthesis were detectable at day 6 and increased over time. Therefore, we demonstrated that HPCs were induced to differentiate into functional mature hepatocytes in vitro, suggesting that factor-treated HPCs may be further explored as a means of liver cell transplantation.

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Hepatic progenitor cells (HPCs) are a potential cell source for liver cell transplantation but do not function like mature liver cells. We sought an effective and reliable method to induce HPC maturation. An immortalized HP14.5 albumin promoter-driven Gaussian luciferase (ALB-GLuc) cell line was established from HPCs isolated from fetal mouse liver of post coitus day 14.5 mice to investigate the effect of induction factors on ALB promoter. HP14.5 parental cells were cultured in DMEM with different combinations of 2% horse serum (HS), 0.1 µM dexamethasone (DEX), 10 ng/mL hepatic growth factor (HGF), and/or 20 ng/mL fibroblast growth factor 4 (FGF4). Trypan blue and crystal violet staining were used to assess cell proliferation with different induction conditions. Expression of hepatic markers was measured by semi-quantitative RT-PCR, Western blot, and immunofluorescence. Glycogen storage and metabolism were detected by periodic acid-Schiff and indocyanine green (ICG) staining. GLuc activity indicated ALB expression. The combination of 2% HS+0.1 µM Dex+10 ng/mL HGF+20 ng/mL FGF4 induced the highest ALB-GLuc activity. Cell proliferation decreased in 2% HS but increased by adding FGF4. Upon induction, and consistent with hepatocyte development, DLK, AFP, and CK19 expression decreased, while ALB, CK18, and UGT1A expression increased. The maturity markers tyrosine aminotransferase and apolipoprotein B were detected at days 3 and 6 post-induction, respectively. ICG uptake and glycogen synthesis were detectable at day 6 and increased over time. Therefore, we demonstrated that HPCs were induced to differentiate into functional mature hepatocytes in vitro, suggesting that factor-treated HPCs may be further explored as a means of liver cell transplantation.

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OBJECTIVE: The aim of this study was to evaluate the potential application of biodegradable nanoparticles (NPs) containing indocyanine green (ICG) in photodynamic therapy (PDT). METHODS: Important parameters, such as particle size and external morphology, were established by dynamic light scattering (DLS) and scanning electron microscopy (SEM). Also, drug encapsulation efficiency and in vitro release behavior were evaluated by spectroscopic methods. RESULTS: The particles are spherical in shape, they exhibit an 817-nm diameter, and they have a low tendency to aggregate. The loading efficiency was 65%. ICG photophysical parameters showed a bathocromic shift in ICG-loaded nanoparticles (ICG-NP). Analysis of the cell P388-D1 in the presence of the ICG-NP by SEM showed that the majority of the nanoparticles were uptaken by phagocytic cells after 2 h of incubation. After laser irradiation photodamage was observed in P388-D1 cells where ICG-NPs had been uptaken by phagocytic cells. CONCLUSION: Polymeric NPs work as an efficient drug delivery system for PDT drugs, and this approach can be used in the administration of amphiphilic photosensitizers in the treatment of neoplasic cells.

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To examine whether hepatic drug metabolism is altered in patients with cystic fibrosis (CF), we evaluated the pharmacokinetics of three model pharmacologic substrates (antipyrine, a marker of hepatic oxidative metabolism; lorazepam, a marker of hepatic glucuronosyltransferase activity; and indocyanine green (ICG), a marker of hepatic blood flow and biliary secretion) in 14 patients with CF (14.6 to 29.2 years of age) and in 12 children and adolescents with cancer (7.2 to 19.4 years of age), which was treated with only surgery and radiation. Each study subject received a single intravenous dose of the combined model substrates (0.03 mg/kg lorazepam, 10 mg/kg antipyrine, and 0.5 mg/kg ICG) for 5 minutes, followed by repeated blood sampling (n = 10) during a 24-hour postinfusion period. Patients with CF had a significantly greater plasma clearance of lorazepam (56.5 +/- 5.2 vs 25.9 +/- 1.9 ml/min/m2) and ICG (892.5 +/- 176.4 vs 256.5 +/- 41.7 ml/min/m2) but not of antipyrine (27.2 +/- 3.8 vs 20.7 +/- 2.0 ml/min/m2) in comparison with control subjects. The apparent steady-state volume of distribution for lorazepam, ICG, and antipyrine was significantly higher in the patients with CF (2.0-, 3.1-, and 1.4-fold, respectively) than in control subjects. Clearance of the model substrates did not correlate with standard biochemical markers of hepatic function. Similarly, no significant relationships were observed between the clearance or steady-state volume of distribution of the compounds and the National Institutes of Health prognostic scores for the patients with CF. These data demonstrate that the plasma clearance of lorazepam and ICG is increased in patients with CF and suggest that hepatic glucuronosyltransferase activity and biliary secretory capacity are enhanced in this disease.

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It is shown that in children suffering from protein calorie malnutrition the mortality associated with fatty liver and liver failure is high. This study was designed to measure functional liver capacity using kinetics analysis of ICG uptake by the liver in animal models of malnutrition and fatty liver disease. Two models were used. Rats were kept either on a 5 percent low protein diet for 6, 12 and 20 days, or on a choline deficient diet for 2 and 6 days. Fatty liver developed in both groups of animals although the changes on the choline deficient diet were more marked than on the protein deficient diet. The choline deficient animals developed a more intense fatty liver at an earlier stage. On the low protein diet the accumulation of fat was inversely correlated to the change in the rate of hepatic uptake of ICG. This relationship could not be demonstrated in the choline deficient group; however, these animals did show a negative correlation between the rate of ICG uptake and FFDLW. It is suggested that the rate of ICG uptake may reflect the rate of liver is discussed in this light (Summary)

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