Hepato-biliary profile of potential candidate liver progenitor cells from healthy rat liver

Figure 1 Rat fibroblastic-like liver derived cell obtention from adult liver parenchymal fraction. A: Cell suspension 24 h after liver parenchyma collagenase A digestion; B: 21 d culture: Rat fibroblastic-like liver derived cells (rFLDC) emergence and hepatocyte death; C: Culture purification due to rFLDC proliferation.

Figure 2 Proliferative capacity of rat fibroblastic-like liver derived cells. Average of cumulative population doubling of rat fibroblastic-like liver derived cells for passage 0 to passage 50.

Figure 3 Representative surface markers expression analysis by flow cytometry. Results are mean ± SE of positive cells for 5 independent experiments.

Figure 4 Rat fibroblastic-like liver derived cells mesenchymal characterization by immunocytochemistry. A-C: Rat bone marrow mesenchymal stem cells (MSC); D-F: Rat fibroblastic-like liver derived cells. ASMA: α-smooth muscle actin.

Figure 5 Representative reverse transcription-polymerase chain reaction characterization of undifferentiated and differentiated rat fibroblastic-like liver derived cells in comparison with rat hepatocyte suspension. Hepatic markers: Cytokeratin 8 (CK8), hepatic nuclear factor 4 (HNF4), tyrosine aminotransferase (TAT), tryptophan 2,3-dioxygenase (TDO), α-fetoprotein (αFP), albumin (Alb), UDP glucuronosyltransferase 1A1 (UGT1A1), glucose-6-phosphatase (G6Pase); Biliary marker: CK19; Mesenchymal markers: Vimentin (Vim), fibronectin (Fn), α-smooth muscle actin (ASMA); Cell surface markers: Integrin β-1 (CD29), hyaluronic acid receptor (CD44), tyrosine phosphatase (CD45), ecto-5’-nucleotidase (CD73), Thy-1 (CD90), endoglin (CD105); Housekeeping gene: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH).

Figure 6 Evaluation of osteogenic and adipogenic differentiation potential of rat fibroblastic-like liver derived cells (alizarin red and red oil O coloration). rFLDC: Rat fibroblastic-like liver derived cells; rBM-MSC: Rat bone marrow-derived mesenchymal stromal cells.

Figure 7 Rat fibroblastic-like liver derived cells hepatogenic differentiation. A: Undifferentiated; B, C: Differentiated.

Figure 8 Hepatic functions. A-C: Glycogen storage ability assessed by periodic acid schiff reaction; D-F: Glucose-6-phosphatase (G6Pase) activity after incubation with glucose-6-phosphate and lead nitrate. rFLDC: Rat fibroblastic-like liver derived cells.

Figure 9 Bilirubin conjugation assay. Comparison between rat hepatocytes, undifferentiated and differentiated rat fibroblastic-like liver derived cells.