Human Liver Cells Expressing Albumin and Mesenchymal Characteristics Give Rise to Insulin-Producing Cells
Figure 4
Isolated cultures of eGFP-positive cells express hepatic markers. Adult human liver cells were infected with the reporter virus and ALB-Cre virus. Two weeks later, eGFP-positive and DsRed2-positive cells were separated using a cell sorter. The eGFP-positive (a–c) and DsRed2-positive (d–f) cells were separately cultured and imaged 24 hours later for eGFP ((a) and (d), green) and DsRed2 ((b) and (e), red) fluorescence. (c and f) eGFP and DsRed2 merge images. Original magnification . Quantitative RT-PCR in eGFP-positive cells compared to DsRed2-positive cells using mRNA extracted 2 weeks after sorting of (g) hepatic (albumin, ADH1b, GLUL, and CEBPβ), (h) stellate cell (SMA, desmin, and GFAP), and (i) mesenchymal stem cell (CD105, CD29. CD90, and CD73) marker expression. The results were normalized to β-actin expression within the same cDNA sample and presented as the relative levels of the mean ± standard deviation of DsRed2-positive versus eGFP-positive cells, from 3 different cultures. *, **. (j) The expression of the mesenchymal marker CD105 was analyzed in the eGFP and (k) DsRed2-positive cells by flow cytometry.
