During the last decade, hepatocyte transplantation has been suggested as a safe and potentially effective clinical option for the treatment of acute or decompensating chronic liver failure as well as for hereditary liver disease. Currently, one of the major limiting factors for clinical application is the insufficient access to suitable liver cell preparations. In cooperation with the German and Catalane organ procurement organizations, a routine procedure for the isolation of hepatocytes from donor organs rejected for transplantation (n = 117) has been established. The process is performed according to the current EC Guidelines for Good Manufacturing Practice (cGMP) and all corresponding national laws and regulations concerning donor organ and tissue procurement. In about 50% of the cases (n = 58) the three-step perfusion procedure has been completed with an average total cell yield of 5.9 × 10 9 cells per organ, the cell preparations displaying a mean viability of 64%. The mean specific yield was 3.6 × 10 6 total and 2.6 × 10 6 viable cells per gram liver tissue, respectively. Specific cell yields from three infantile donor livers were considerably higher. No correlation between isolation efficiency and cold ischemia time or donor age was found within the adult organ donors. In contrast, organs with a severe steatosis generally did not result in successful cell isolation. Results of sterility and endotoxin determination are also presented. In summary, a standardized and cGMP conform method of hepatocyte isolation from nontransplantable liver organs was established, which reproducibly yields large amounts of hepatocytes suitable for therapeutic application.
Corneal opacity, reversible retinal lipidosis and irreversible receptor cell degeneration are known to occur after long-term treatment with chloroquine. Female albino Wistar rats (initial age 6 weeks, weight between 100 and 150 g) were treated orally with chloroquine (40-60 mg/kg body weight) for 4 weeks and with 70–80 mg/kg body weight for the following 4 (group A) and 8 weeks (group B). The animals were submitted to electroretinography ERG, and the retinas were prepared for histological investigation. After treatment with chloroquine for 8 weeks, lipidosis-like inclusions could be seen in the rat retina. A deformation of the receptor cell layer was not observed by light microscopy. The a-wave amplitude decreased to 33% and the b-wave amplitude to 40% of the values before treatment. In contrast to group A, we found receptor cell degeneration and macrophage-like cells in the peripheral and central retina in rats treated for 12 weeks. These changes were probably responsible for a-wave and b-wave reductions of 50 and 79% of values before treatment, respectively. It can be assumed that changes in ERG parameters in the first period are caused by lipidosis. Later extremer deformation is induced by receptor cell degeneration and accompanying lipidosis.
Seen from the point of view of function, it is doubtful whether lipidosis is the primary cause of changes in the electroretinogram or of receptor cell degeneration.
Chronic administration of the cationic amphiphilic anorexigenic drug chlorphentermine to rats has previously been shown to induce extraocular and ocular lipidosis: large numbers of lipidosis-related cytoplasmic inclusions can be found in the pigment epithelium and smaller numbers in the neuroretina. In the present study, female albino Wistar rats were treated orally with chlorphentermine (30-45 mg/kg body weight) for 4-16 weeks. The animals were submitted to electroretinography, and the retinae were prepared for histological investigations. Our histological findings corresponded to previous reports. The changes in electroretinographic parameters were low. The clearest change was a reduction of the b-wave amplitude of 20% after 12 and 16 weeks of treatment compared with the values before drug treatment. The a-wave amplitude did not differ from that in the control group. Lipidosis in the neuroretina may be the reason for functional influences on the b-wave amplitude. The function of the receptor cells, which is represented by the a-wave, appeared unaffected by chlorphentermine.
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