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1.
Figure 4

Figure 4. Liver-Derived extracellular matrix promotes differentiation of hAECs into hepatic cells over time. From: Hepatic Differentiation of Amniotic Epithelial Cells.

Gene expression of hAECs over a three week differentiation protocol on L-ECM. mRNA levels expressed as arbitrary numbers normalized to Cyclophilin-A.

Fabio Marongiu, et al. Hepatology. ;53(5):1719-1729.
2.
Figure 3

Figure 3. Basement membrane matrix proteins influence hepatic differentiation of hAECs. From: Hepatic Differentiation of Amniotic Epithelial Cells.

Gene expression of hAECs after two week of differentiation. hAECs were culture on Matrigel or liver-derived ECM (L-ECM). mRNA levels expressed as arbitrary numbers normalized to Cyclophilin-A.

Fabio Marongiu, et al. Hepatology. ;53(5):1719-1729.
3.
Figure 6

Figure 6. Naïve hAECs differentiate into mature hepatocytes upon transplantation into SCID/beige mouse liver. From: Hepatic Differentiation of Amniotic Epithelial Cells.

Gene expression of hAECs 6 months after transplantation into mouse host livers. Comparison with undifferentiatied hAECs, human fetal liver and human adult liver. mRNA levels detected with human specific primer/probes and expressed as arbitrary numbers normalized to Cyclophilin-A.

Fabio Marongiu, et al. Hepatology. ;53(5):1719-1729.
4.
Figure 1

Figure 1. Activin-A pretreatment is not required for hepatic differentiation of hAECs. From: Hepatic Differentiation of Amniotic Epithelial Cells.

Gene expression of hAECs after a 4 day treatment in the presence or absence of 100ng/ml Activin-A. mRNA levels are expressed as arbitrary numbers normalized to Cyclophilin-A and relative to freshly isolated cells. *P < 0.05; **P < 0.005.

Fabio Marongiu, et al. Hepatology. ;53(5):1719-1729.
5.
Figure 2

Figure 2. Co-culture of hAECs with Mouse hepatocytes improves hepatic differentiation of hAECs. From: Hepatic Differentiation of Amniotic Epithelial Cells.

(A) Gene expression of hAECs after co-culture with mouse hepatocytes (mHeps) at day 16. mRNA levels expressed as arbitrary numbers normalized to Cyclophilin-A and relative to mRNA levels at day 1. (B) Testosterone metabolism of hAECs after co-culture with mHeps at day 20, after 3-days induction with Rifampicin and Phenobarbital. Results measured by HPLC and expressed as 6β-hydroxytestosterone metabolite formation, relative to mHep control sample. *P < 0.05; **P < 0.005; #P < 0.001.

Fabio Marongiu, et al. Hepatology. ;53(5):1719-1729.
6.
Figure 7

Figure 7. Naïve rAECs integrate and form clusters of mature hepatocytes upon transplantation into syngenic rats. From: Hepatic Differentiation of Amniotic Epithelial Cells.

Immunofluorescence staining of serial frozen section of rat livers after transplantation of rAECs. Recipient animals were DPPIV while transplanted rAECs were isolated from DPPIV+ tissues. Clusters of positive differentiated cells can be found into the host liver. (A) Double stain for DPPIV (green) and CYP2E1 (red). (B) Double stain for DPPIV (green) and CYP3A1 (red). (C) Double stain for DPPIV (red) and Albumin (green). Differentiated rAECs expressed hepatocyte markers at levels comparable to the surrounding liver.

Fabio Marongiu, et al. Hepatology. ;53(5):1719-1729.
7.
Figure 5

Figure 5. Liver-Derived extracellular matrix efficiently promotes differentiation of hAECs into hepatic cells with metabolic activity and inducible enzymes. From: Hepatic Differentiation of Amniotic Epithelial Cells.

(A) percent of ammonia metabolized by differentiated hAECs in a time period of 3 and 6 hours; (B) LC-MS chromatograms of 17-OHPC and its metabolites derived from incubation of 17-OHPC with differentiated hAECs and fresh human fetal hepatocytes. Incubation of 17-OHPC with differentiated hAECs generated 4 metabolites of which 2 were the major metabolites (M2 and M4). Incubation of 17-OHPC with human fetal hepatocytes generated numerous metabolites of which M1, M2 and M4 were common with differentiated hAECs. (C) Gene expression levels of hAEC-derived hepatic cells after 3-days induction with Phenobarbital (PB) or β-naphtoflavone (BNF). mRNA levels expressed as arbitrary numbers normalized to Cyclophilin-A and relative to untreated control (DMSO).

Fabio Marongiu, et al. Hepatology. ;53(5):1719-1729.

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