Fig. 6.
Experimental design to study the effects of alpha-1 antitrypsin on hepatocyte engraftment. Primary rat hepatocytes were isolated from either Sprague Dawley wild-type (WT) rats or eGFP Wistar rats and cryopreserved. WT cells were labelled with the cell tracking dye CM-DiL. 15 × 106 hepatocytes were transplanted intrasplenically following the IV injection of 120 mg/kg AAT or water for injection (controls). Animals that received CM-DiL-labelled cells were sacrificed at 48 h and 1 week and engraftment quantified in the entire liver using the IVIS imaging machine. Further analysis was carried out in frozen liver sections by co-staining with DAPI and CD68 to ensure macrophages that engulfed transplanted hepatocytes were not included in the analysis. Analysis was further confirmed by PCR analysis for the Y chromosome (SRY). For long-term engraftment, rats received eGFP hepatocytes and engraftment was quantified in paraffin sections by co-labelling with DAPI