Therapeutic efficiency of human amniotic epithelial stem cell-derived functional hepatocyte-like cells in mice with acute hepatic failure

Abstract

BackgroundHepatocyte transplantation has been proposed as an effective treatment for patients with acute liver failure (ALF), but its application is limited by a severe shortage of donor livers. Human pluripotent stem cells (hPSCs) have emerged as a potential cell source for regenerative medicine. Human amniotic epithelial stem cells (hAESCs) derived from amniotic membrane have multilineage differentiation potential which makes them suitable for possible application in hepatocyte regeneration and ALF treatment.MethodsThe pluripotent characteristics, immunogenicity, and tumorigenicity of hAESCs were studied by various methods. hAESCs were differentiated to hepatocyte-like cells (HLCs) using a non-transgenic and three-step induction protocol. ALB secretion, urea production, periodic acid-Schiff staining, and ICG uptake were performed to investigate the function of HLCs. The HLCs were transplanted into ALF NOD-SCID (nonobese diabetic severe combined immunodeficient) mouse, and the therapeutic effects were determined via liver function test, histopathology, and survival rate analysis. The ability of HLCs to engraft the damaged liver was evaluated by detecting the presence of GFP-positive cells.ResultshAESCs expressed various markers of embryonic stem cells, epithelial stem cells, and mesenchymal stem cells and have low immunogenicity and no tumorigenicity. hAESC-derived hepatocytes possess the similar functions of human primary hepatocytes (hPH) such as producing urea, secreting ALB, uptaking ICG, storing glycogen, and expressing CYP enzymes. HLC transplantation via the tail vein could engraft in live parenchymal, improve the liver function, and protect hepatic injury from CCl4-induced ALF in mice. More importantly, HLC transplantation was able to significantly prolong the survival of ALF mouse.ConclusionWe have established a rapid and efficient differentiation protocol that is able to successfully generate ample functional HLCs from hAESCs, in which the liver injuries and death rate of CCl4-induced ALF mouse can be significantly rescued by HLC transplantation. Therefore, our results may offer a superior approach for treating ALF.

Transcriptomic and computational analysis identified LPA metabolism, KLHL14 and KCNE3 as novel regulators of Epithelial-Mesenchymal Transition

“…Indeed, AEC have been proved capable to interact with different typologies of host damaged tissue such as lung 10 , liver 11 , cholangiocyte 12 , CNS 13 , tendon 14 or sinusoidal endothelial cells 11 by promoting regeneration, managing inflammation and replacing damaged cells with functional/mature ones (without cell fusion). The level of maturation implies either a tissue replacement as epithelial cells [15][16][17][18][19][20][21] , or as mesenchymal cells by experiencing 1 Faculty of Bioscience and Technology for Food, Agriculture and Environment, University of Teramo, Teramo, Italy.…”

Alternative Cell Sources for Liver Parenchyma Repopulation: Where Do We Stand?

“…Generally, HLCs derived from MSCs possess some mature and fetal hepatocyte features as has been shown by transcriptome and functional studies [102,103]. It should be noted that some differences are also reported depending on the origin of the MSC population (e.g., BM vs. umbilical cord vs. liver-derived MSCs; pediatric vs. adult MSCs) [87,95,104].…”

“…For instance, Liu et al used a 14-day cytokine-based differentiation protocol to obtain HLCs from human AECs in 2D culture. Following grafting 2 × 10 6 of such AEC-HLCs in CCl 4 treated NOD-SCID mice, human cells were detected in the recipient liver (although the RI was not quantified) and liver injury was attenuated [104]. However, it remains to be addressed whether naïve AECs or in vitro differentiated AEC-HLCs are superior as an alternative cell source for alleviating liver failure and/or repopulating the host liver [114].…”

Differentiation of Cells Isolated from Afterbirth Tissues into Hepatocyte-Like Cells and Their Potential Clinical Application in Liver Regeneration

“…However, such concerns are not raised by the use of amnion epithelial cells [11] and, as it can be assumed, HLC obtained from them. Preclinical studies on mice [14,113] using HLC obtained from hAEC have shown, that their use in the treatment of liver failure in humans does not require the use of immunosuppressive treatment [13].…”