Based on their ability to proliferate and the capacity to differentiate into specific cell types, hepatic progenitor/stem cells (HPCs) from adult human liver may have potential therapeutic effects on end-stage liver failure. In addition, adult HPCs have a reduced risk of teratoma formation and are not subject to the same ethical issues as fetal HPCs or embryonic stem cells 1. The HPCs from rhesus monkeys are relevant because they may serve as a valuable preclinical model for assessment of cell therapy in humans. To date, there are no reports of HPCs or liver epithelial progenitor cells (LEPCs) isolated from normal adult rhesus monkey although a few studies in other species were reported 2, 3. We report here for the first time the successful isolation of rhesus monkey LEPCs (mLEPCs) from normal adult livers (n = 12).

The polygonal epithelial cells from single aggregates proliferated rapidly and formed moderate colonies within 4-5 days (Supplementary information, Figure S2A) and large colonies within 8–10 days (Supplementary information, Figure S2B) consisting of thousands of cells with similar morphology to previously identified HPCs 5, 6. Almost all of the polygonal epithelial cells in different colonies were positive for E-cad (Figure 1B-i), a marker of HPCs 7, 8. FACS sorting was then employed to yield E-cad+CD34–CD45–CD90– cells (Supplementary information, Figure S3A-S3C), and the E-cad+ cells were enriched from 96.3% (Supplementary information, Figure S3C) to 99.8% after sorting (Figure 1C). The sorted cells rapidly formed colonies (Supplementary information, Figure S2C), stained positively for hepatic epithelial markers CK7 (Figure 1B-ii) as well as CK8/CK18 (Figure 1B-iii), and exhibited a hepatobiliary phenotype of HPCs 9: expression of hepatocytic (albumin, APOH) and cholangiocytic (IB4, CX43) genes as well as HNF1β and HNF3β (Figure 1B-iv). More importantly, the cells were cultured for at least 20 passages without change of expression profiles and cell morphology (data not shown). All of the above results indicated that the FACS-sorted cells might be HPCs.

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