Research Article

Immunoprofiling of Adult-Derived Human Liver Stem/Progenitor Cells: Impact of Hepatogenic Differentiation and Inflammation

Figure 3

Downregulation of CD200 expression after in vitro hepatogenic differentiation of ADHLSC. ADHLSCs were submitted to an in vitro hepatogenic differentiation protocol and processed for CD200 expression analysis. (a) Total RNA extracted from both undifferentiated and differentiated MSCs (the umbilical cord MSC (UCMSC), bone marrow MSC (BM-MSC), and adipose tissue MSC (AT-MSC)) was retrotranscribed, the cDNA synthesized, and RT-qPCR applied for CD200 gene marker. Hepatogenic differentiation was correlated with a strong downregulation of this marker. (b) Total proteins extracted from differentiated and undifferentiated ADHLSCs and UCMSC were analyzed using western blotting. Hepatogenic differentiation was associated with a significant decrease of CD200 protein expression. (c) Downregulation of CD200 mRNA occurs early at the first step of the hepatogenic differentiation protocol as demonstrated using RT-qPCR. (d) Downregulation of CD200 expression in differentiated ADHLSCs occurs under EGF and oncostatin M treatment. ADHLSCs were treated with one or the other growth factor/cytokine used in the hepatogenic differentiation protocol (epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), or oncostatin M (OSM)). Cells were treated for 3 days using the same basal medium as for differentiation experiments and under serum-free conditions. Total RNA for each group was extracted and retrotranscribed for CD200 mRNA analysis using RT-qPCR. Data shown represent the mean ± SEM of three different experiments as indicated in the graph (; versus corresponding untreated cells, unpaired t-test).