By R.H. Burdon and P.H. van Knippenberg (Eds.)
This ebook units out essentially and successfully the education and dealing tools of laboratory concepts concerning remoted hepatocytes and should make lifestyles more straightforward for each laboratory employee desirous about those concepts.
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Additional resources for Isolated Hepatocytes Preparation, Properties and Applications
The perfusate exits the liver via the hepatic veins, which empty into the inferior vena cava. A length of tubing, connected to the outflow cannula, drains fluid from the vena cava back into the reservoir. Efflux from the liver is assisted by a siphoning effect, achieved by mounting the rat on a platform some 15 cm above the reservoir (Fig. 4). The apparatus should be kept clean and free of bacterial growth. After each perfusion it is rinsed thoroughly with deionized water, and Ch. 2. HIGH-YIELD PREPARATION H p L BT RES GI GO 0 T F 4 heaters Pump lung bubbletrap resetvoir gasinlet gasoutlet on-off tap tray filter direction of flow 23 1.
4). Washing removes constituents that have leaked from intact or damaged cells, cell debris and a substantial proportion of the damaged cells, as well as almost all non-parenchymal 32 ISOLATED HEPATOCYTES cells. Higher centrifugal forces result in slightly larger yields of hepatocytes but more contamination with damaged hepatocytes and non-parenchymal cells. Liver perfusion requires a certain amount of practice. Not infrequently the beginner encounters problems with placement of cannulae, so that swelling of the organ occurs, or alternatively, the portal vein is not successfully cannulated, or the cannula slips out of the vein during the perfusion.
It can be inferred, therefore, that during the first stage of the twostage procedure introduced by Seglen (1972, 1976), pre-perfusion with a medium lacking Ca2+brings about cleavage of the desmosomes to a stage of irreversibility. In consequence, subsequent perfusion of the liver with a medium rich in Ca2+is without effect on cell adhesion. Ch. 1. 2). The role of Ca2+in cell separation is seen to be complex. These ions are known to be essential for collagenase activity, so that the inclusion of any Ca2+-bindingagent in the medium will prevent collagenase action.
Isolated Hepatocytes Preparation, Properties and Applications by R.H. Burdon and P.H. van Knippenberg (Eds.)