High-efficiency c-Myc-mediated induction of functional hepatoblasts from the human umbilical cord mesenchymal stem cells

Abstract

Background Direct reprogramming of human fibroblasts to hepatocyte-like cells was proposed to generate large-scale functional hepatocytes demanded by liver tissue engineering. However, the difficulty in obtaining large quantities of human fibroblasts greatly restricted the extensive implementation of this approach. Meanwhile, human umbilical cord mesenchymal stem cells (HUMSCs) are the preferred cell source for HLCs with the advantages of limited ethical concerns, easy accessibility, and propagation in vitro. However, no direct reprogramming protocol for converting HUMSCs to hepatoblast-like cells (HLCs) has been reported. Methods HLCs were successfully generated from HUMSCs by forced expression of FOXA3, HNF1A, and HNF4A (collectively as 3TFs) and c-Myc. In vitro and in vivo functional experiments were conducted to demonstrate the hepatic phenotype, characterization, and function of HUMSC-derived HLCs (HUMSC-iHeps). ChIP-seq and RNA-seq were integrated to reveal the potential molecular mechanisms underlying c-Myc-mediated reprogramming. Results We showed that c-Myc greatly improved the trans-differentiation efficiency for HLCs from HUMSCs, which remained highly efficient in reprogramming fibroblasts into HLCs, suggesting c-Myc could promote direct reprogramming and its potentially widespread applicability for generating large amounts of HLCs in vitro. Mice transplantation experiments further confirmed the therapeutic potential of HUMSC-iHeps by liver function restoration and survival prolongation. Besides, in vivo safety assessment demonstrated the low risk of the tumorigenic potential of HUMSC-iHeps. We found that c-Myc functioned predominantly at an early phase of reprogramming, and we further unraveled the regulatory network altered by c-Myc. Conclusions c-Myc enhanced reprogramming efficiency of HLCs from HUMSCs. A large scale of functional HLCs generated more conveniently from HUMSCs could benefit biomedical studies and applications of liver diseases.

Isolation of Homogenous Population of Human Amniotic Epithelial Stems Cells

“…In other studies, they found specific enzymatic digestion of amniotic membrane in which human amniotic epithelial stem cells (hAESCs) and amniotic mesenchymal stem cells (hAMSCs) were digested by protease enzymes trypsin and collagenases respectively 15,19,23,28 . Human amniotic epithelial stem cells are in a single layer bound with weak cellular interaction so trypsin dissociates them easily 20,27 .…”

Integrative analysis reveals a clinicogenomic landscape associated with liver metastasis and poor prognosis in hepatoid adenocarcinoma of the stomach

“…As a famous oncogene at 8q24.21 region, MYC was identified as one of the core genes regulating stem cell-like properties of cancer cells 68 . A recent study illustrated that high-efficiency c-Myc enhanced the conversion from mesenchymal stem cells to hepatoblast-like cells 69 . In this study, we found that MYC and its regulators ( HUWE1 and MYCBP2 ) were enriched in the signaling pathways regulating pluripotency of stem cells (Figure 4 A-B).…”

Updated Perspectives on Direct Vascular Cellular Reprogramming and Their Potential Applications in Tissue Engineered Vascular Grafts

“…Studies should look for multi-year assessments of tumorigenicity in longer-living animals (e.g., pigs and sheep) and closer animal relatives to humans. Furthermore, there is an inherent tumour risk with using pluripotency factors, notably c-MYC (in OSKM), which has been shown as both dispensable and a facilitator of lineage-conversion [ 109 , 110 ]. As further research occurs, we may find efficient non-oncogenic factors for direct cellular reprogramming.…”