Objective To acquire oval cells (progenitor stem cells) from adult rat liver of different models including diabetic rats. Methods Thirty Sprague-Dawley (SD) rats were divided into 5 groups randomly: control, 2-acetyla...Objective To acquire oval cells (progenitor stem cells) from adult rat liver of different models including diabetic rats. Methods Thirty Sprague-Dawley (SD) rats were divided into 5 groups randomly: control, 2-acetylaminofluorene (2-AAF), 2-AAF+partial hepatectomy (PH), 2-AAF+carbon tetrachloride (CCl4), and diabetic groups. As two-step collagenase perfusion protocol of Seglen, oval cells were isolated by Percoll density gradient centrifugation. Thy1.1 positive cells were sorted by flow cytometry, and then cultured in Dulbecco's minimum Eagle's medium (DMEM). Immunofluorescence staining was applied to labelling Thy1.1. Results Different rates of Thy1.1 positive oval cells were found in different rat model groups: 0.5% in 2-AAF, 0.3% in 2-AAF+PH, 0.2% in 2-AAF+CCl4 , 0.1% in diabetic, and 0.0% in control. Isolated cells adhered to plate with fusiform or polygon as epithelial cells. Conclusion Progenitor stem cells exist in injured liver tissue including those from diabetic rats.展开更多
基金Supported by Shanghai Municipal Education Commission Key Project (07ZZ35)
文摘Objective To acquire oval cells (progenitor stem cells) from adult rat liver of different models including diabetic rats. Methods Thirty Sprague-Dawley (SD) rats were divided into 5 groups randomly: control, 2-acetylaminofluorene (2-AAF), 2-AAF+partial hepatectomy (PH), 2-AAF+carbon tetrachloride (CCl4), and diabetic groups. As two-step collagenase perfusion protocol of Seglen, oval cells were isolated by Percoll density gradient centrifugation. Thy1.1 positive cells were sorted by flow cytometry, and then cultured in Dulbecco's minimum Eagle's medium (DMEM). Immunofluorescence staining was applied to labelling Thy1.1. Results Different rates of Thy1.1 positive oval cells were found in different rat model groups: 0.5% in 2-AAF, 0.3% in 2-AAF+PH, 0.2% in 2-AAF+CCl4 , 0.1% in diabetic, and 0.0% in control. Isolated cells adhered to plate with fusiform or polygon as epithelial cells. Conclusion Progenitor stem cells exist in injured liver tissue including those from diabetic rats.
