Isolation of EpCAM+/CD133- Hepatic Progenitor Cells

Abstract

Progenitor cell-derived hepatocytes are critical for hepatocyte replenishment. Therefore, we established a line of human hepatic progenitor (HNK1) cells and determined their biological characteristics for experimental and therapeutic applications. HNK1 cells, isolated from human noncirrhotic liver samples with septal fibrosis, showed high expression of the hepatic progenitor cell (HPC) markers EpCAM, CK7, CK19, alpha-fetoprotein (AFP), CD90 (Thy1), and EFNA1. Expression of CD133 was very low. Ductular reactions at the periphery of cirrhotic nodules were immunohistochemically positive for these HPC markers, including EFNA1. Sodium butyrate, a differentiation inducer, induced hepatocyte-like morphological changes in HNK1 cells. It resulted in down-regulation of the hepatic progenitor cell markers EpCAM, CK7, CK19, AFP, and EFNA1 and up-regulation of mature hepatocyte markers, including albumin, CK8, and CK18. Furthermore, sodium butyrate treatment and a serial passage of HNK1 cells resulted in enhanced albumin secretion, ureagenesis, and CYP enzyme activity, all of which are indicators of differentiation in hepatocytes. However, HNK1 cells at passage 50 did not exhibit anchorage-independent growth capability and caused no tumors in immunodeficient mice, suggesting that they had no spontaneous malignant transformation ability. From this evidence, HNK1 cells were found to be EpCAM(+)/CD133(-) hepatic progenitor cells without spontaneous malignant transformation ability. We therefore conclude that HNK1 cells could be useful for experimental and therapeutic applications.

Inhibition of notch signaling pathway prevents cholestatic liver fibrosis by decreasing the differentiation of hepatic progenitor cells into cholangiocytes

“…23 Sox9 is an endodermal transcription factor, and EpCAM is a stem/progenitor cell surface marker. [24][25][26] Immunostaining showed that CK19 and OV6 were expressed in proliferated cholangiocytes after BDL treatment (Figures 2a and b). Co-immunostaining revealed that OV6/CK19, Sox9/CK19 were extensively co-expressed in proliferated cholangiocytes (Figures 2c and d) and immunoblotting confirmed CK19, OV6, Sox9, and EpCAM protein gradually increased (Figure 2e).…”

Inhibition of the Notch Signaling Pathway Reduces the Differentiation of Hepatic Progenitor Cells into Cholangiocytes in Biliary Atresia

“…SB is a commonly used reagent for inducing cholangiocyte differentiation [25]. CK19 is a marker of bile epithelial cells [26], Sox9 is an endodermal transcription factor [27], and EpCAM is a stem/progenitor cell surface marker [28]. When WB-F344 cells were treated with SB, the mRNA and protein levels of CK19, Sox9, and EpCAM were dramatically increased in the SB group compared with the control group, which was reduced by DAPT treatment (Fig.…”

Specific Markers for Hepatic Progenitor Cells

“…Another study suggested that HPCs express the same marker as primitive bile duct cells of the ductal plate, AFP, but have different phenotypic features from cholangiocytes and hepatocytes. HPCs characteristic were in the middle between those of stem cells and mature cells (Van Den Heuvel et al, 2001;Petersen et al, 1998;Li et al, 2014;Hao et al, 2013). This finding suggested that HPCs have bipotential capacity to differentiate into both cholangiocytes and hepatocytes.…”

“…Hao et al (2013) found expression of EFNA1 in ductular reaction is correlated to AFP. EFNA1 expressions were stronger than AFP, thus EFNA1 can be used as HPCs.…”

“…HPCs also express cholangiocytes markers (CK19) moderately and HSCs marker (CD45 and CD109) strongly. Akiba et al (2013) CD56, c-kit (CD117), CD133, EpCAM Badrawy et al (2013) OV6, AFP Chen et al (2015) OV6, CK19 Hao et al (2013) EpCAM, CD133, AFP, EFNA1 Jia et al (2013) OV6 Li et al (2014) Alb, AFP, CK8, CK18, CK19, CD90, c-kit (CD117), OV6, CD45, CD109 Libbrecht et al (2000) CK7, CK19, CK8, CK18, Chrom-A, OV6 Petersen et al (1998) AFP, GGT, OV6, CK19, OC2, Thy-1 Van Den Heuvel et al (2001) OV6, CK19 Ye et al (2014) EpCAM, OV6, CD133, c-kit (CD117) Yun et al (2013) CK19, Oct-3/4, EpCAM, DLK1 Yovchev et al (2007) EpCAM, CD44, CD24, CD133 In addition, Ye et al (2014) used four markers for HPCs (EpCAM, OV6, CD133 and c-kit) and found that HPCs express HSCs marker (c-kit and CD133) and express biliary marker (EpCAM). Another HSCs marker used to identify HPCs was Thy-1 (Petersen et al, 1998).…”