Differential response to hepatic differentiation stimuli of amniotic epithelial cells isolated from four regions of the amniotic membrane

Abstract

Human Amniotic Epithelial Cells (hAEC) isolated from term placenta are a promising source for regenerative medicine. However, it has long been debated whether the hAEC population consists of heterogeneous or homogeneous cells. In a previous study, we investigated the characteristics of hAEC isolated from four different regions of the amniotic membrane finding significant heterogeneity. The aim of this study was to evaluate the hepatic differentiation capability of hAEC isolated from these four regions. Human term placentae were collected after caesarean section and hAEC were isolated from four regions of the amniotic membrane (R1‐R4, according to their relative distance from the umbilical cord) and treated in hepatic differentiation conditions for 14 days. hAEC‐derived hepatocyte‐like cells showed marked differences in the expression of hepatic markers: R4 showed higher levels of Albumin and Hepatocyte Nuclear Factor (HNF) 4α whereas R1 expressed higher Cytochrome P450 enzymes, both at the gene and protein level. These preliminary results suggest that hAEC isolated from R1 and R4 of the amniotic membrane are more prone to hepatic differentiation. Therefore, the use of hAEC from a specific region of the amniotic membrane should be taken into consideration as it could have an impact on the outcome of therapeutic applications.

In Vitro Differentiation of Human Amniotic Epithelial Cells into Hepatocyte-like Cells

“…The process of hepatocyte differentiation is favoured by culturing in extracellular matrix-covered dishes. Substrates tested so far, including porcine-liver-derived extracellular matrix (L-ECM) [ 29 ], decellularized human liver extracellular matrix (hDLM) [ 42 ], laminins 411 (LN-411) and 521 (LN-521) [ 43 ], and the widely used Matrigel (Geltrex), i.e., the cell matrix derived from the Engelbreth-Holm-Swarm tumour [ 16 , 29 , 43 , 44 , 45 ], are characterised by varying effectiveness of differentiation. Some of them, e.g., LN-521, LN-411, and L-ECM, more efficiently increase the expression of genes encoding proteins from the cytochrome P450 family involved in the metabolism of endogenous compounds and xenobiotics [ 29 , 43 ].…”

“…Substrates tested so far, including porcine-liver-derived extracellular matrix (L-ECM) [ 29 ], decellularized human liver extracellular matrix (hDLM) [ 42 ], laminins 411 (LN-411) and 521 (LN-521) [ 43 ], and the widely used Matrigel (Geltrex), i.e., the cell matrix derived from the Engelbreth-Holm-Swarm tumour [ 16 , 29 , 43 , 44 , 45 ], are characterised by varying effectiveness of differentiation. Some of them, e.g., LN-521, LN-411, and L-ECM, more efficiently increase the expression of genes encoding proteins from the cytochrome P450 family involved in the metabolism of endogenous compounds and xenobiotics [ 29 , 43 ]. Matrigel, on the other hand, is the most effective commercially available substrate, promoting the differentiation of stem cells into hepatocytes within 14 days, more than 90% of which are capable of albumin synthesis [ 15 , 45 ].…”

Sub-Regional Differences of the Human Amniotic Membrane and Their Potential Impact on Tissue Regeneration Application

“…Furthermore, in some studies the placental amnion was sub-divided into an area surrounding the umbilical cord (R1, central area) [ Figure 1A (1a)] and an intermediate area (R2) [ Figure 1A (1b)] between R1 and R3, where R3 (peripheral area) [ Figure 1A (1c)] is close to the edge of the placental amnion ( Centurione et al, 2018 ; Passaretta et al, 2020 ).…”

“…They found high levels of alpha-fetoprotein in the central area and the peripheral area of hAM tissue, which could point to a higher hepatic differentiation capacity of this area ( Centurione et al, 2018 ). However, in isolated hAECs upon hepatic differentiation, higher levels of albumin and hepatocyte nuclear factor 4α in the reflected amnion, and higher cytochrome p450 in the central area were found ( Passaretta et al, 2020 ). These results lead to the speculation that hAECs from the central and the reflected area have a higher hepatic differentiation capacity ( Passaretta et al, 2020 ).…”

“…However, in isolated hAECs upon hepatic differentiation, higher levels of albumin and hepatocyte nuclear factor 4α in the reflected amnion, and higher cytochrome p450 in the central area were found ( Passaretta et al, 2020 ). These results lead to the speculation that hAECs from the central and the reflected area have a higher hepatic differentiation capacity ( Passaretta et al, 2020 ).…”