Clonal expansion of hepatic progenitor cells and differentiation into hepatocyte‐like cells

Abstract

Hepatic progenitor cells (HPCs) in adult liver are promising for treatment of liver diseases. A biliary‐derived HPC population in adult mice has been characterized by co‐expression of stem cell marker Sry (sex determining region Y)‐box 9 (SOX9) and biliary marker cytokeratin 7 (CK7). However, isolation of these HPCs in adult healthy liver without any selection procedures remains a big challenge in this field. Here, by establishing a simple and efficient method to isolate and expand the CK7+SOX9+ HPCs in vitro as clones, we acquired a stable and largely scalable cell source. The CK7+SOX9+ progenitor cells were then further induced to differentiate into hepatocyte‐like cells with expression of mature hepatocyte markers albumin (Alb) and hepatocyte nuclear factor 4 alpha (HNF4α), both in vitro and in vivo in the presence of hepatocyte growth factor (HGF) and fibroblast growth factor 9 (FGF9). Furthermore, we found that the HPCs are highly responsive to transforming growth factor‐beta (TGF‐β) signals. Collectively, we identified and harvested a CK7+SOX9+ progenitor cell population from adult mouse liver by a simple and efficient approach. The exploration of this HPC population offers an alternative strategy of generating hepatocyte‐like cells for cell‐based therapies of acute and chronic liver disorders.

Small-molecule inhibitor cocktail promotes the proliferation of pre-existing liver progenitor cells

“…Thus, transplanting LPCs seems to be a more reasonable option. The methods available currently for the purification of LPCs are mostly based on fluorescence-activated cell sorting (FACS) via cell labeling with specific antibodies ( Liu et al., 2019b ; Suzuki et al., 2008 ); however, for clinical application, the safety of antibody-conjugated cell transplantation is also of concern. Therefore, the generation of abundant and clinically available LPCs is a new challenge, and more feasible methods should be developed to attain this goal.…”

Extracellular microparticles derived from hepatic progenitor cells deliver a death signal to hepatoma-initiating cells

“…6 weeks of oral DEN treatment showed the obvious HPC activation (Additional file 2 : Fig. S3), HPCs were labeled with an antibody against CK7 [ 23 ], and in our previous studies, we found that the activation and malignant transformation of HPCs promote hepatocarcinogenesis and HCC recurrence [ 9 – 12 ]. Thus, the rats were intrasplenically administered with 40 µg of apoLTC-MPs, apoHep-MPs, apoHPC-MPs or saline after 6 weeks of oral DEN treatment.…”

“…4 C, D). To corroborate these observations, we also evaluated the expression of the HPC marker SOX9 and CK7 in liver sections by IHC [ 23 , 24 ]. ApoLTC-MPs and the control group showed intense and diffuse positive SOX9 and CK7 staining, while apoHep-MPs caused a small decrease in the SOX9 and CK7 signal and apoHPC-MPs caused a significant decrease in SOX9 and CK7 staining (Fig.…”

Induction and Maturation of Hepatocyte-Like Cells In Vitro: Focus on Technological Advances and Challenges

“…These cells can be lineage-restricted into hepatocytes under different condition, which indicates that they are safe for use in transplantation in vivo (Cardinale et al, 2011;Turner et al, 2011). However, the extraction and separation of HpSCs or hepatoblasts presents a challenge due to the scant numbers of HpSCs (0.5-2.5% of liver parenchyma of all donor ages) and hepatoblasts (<0.01% in adult livers) (Turner et al, 2011;Liu et al, 2019). Although it is possible to obtain proliferative hepatoblasts by transferring both stem maintaining genes and liver specific genes, the final differentiation efficiency seems to be dissatisfactory (only 56.7%) (Yu et al, 2013;.…”