Hepatic Stem/progenitor cells (HSPCs) have gained a large amount of interest for treating acute liver disease. However, the isolation and identification of HSPCs are unclear due to the lack of cell-specific surface markers. To isolate adult HSPCs, we used cell surface-marking antibodies, including CD49f and Sca-1. Two subsets of putative HSPCs, Lin⁻CD45⁻Sca-1⁻CD49f⁺ (CD49f⁺) and Lin⁻CD45⁻Sca-1⁺CD49f⁻ (Sca-1⁺) cells, were isolated from adult mice liver by flow cytometry. Robust proliferative activity and clonogenic activity were found in both CD49f⁺ and Sca-1⁺ cells through colony-forming tests and cell cycle analyses. Immunofluorescence staining revealed that CD49f⁺ cells expressed ALB and CK-19 while Sca-1⁺ cells expressed only ALB, indicating that CD49f⁺ cells were bipotential and capable of differentiating into hepatocyte and cholangiocyte. Consequently, PAS stain showed that differentiated CD49f⁺ and Sca-1⁺ cells synthesised glycogen, indicating they could differentiate into functional hepatocytes. mRNA expression profile indicated that both CD49f⁺ and Sca-1⁺ cells showed differential expression of genes that are associated with liver progenitor function such as Sox9 and EpCam. Moreover, two subsets of putative HSPCs were activated by DDC and we found that their abundance and proliferation increased with age. In summary, we hypothesized that CD49f⁺ cells were a type of potential HSPCs and may be utilised for clinical stem cell therapy.

肝干细胞/祖细胞 (HSPC) 在治疗急性肝病方面引起了极大的兴趣。然而，由于缺乏细胞特异性表面标志物，HSPCs 的分离和鉴定尚不清楚。为了分离成人 HSPC，我们使用了细胞表面标记抗体，包括 CD49f 和 Sca-1。通过流式细胞术从成年小鼠肝脏中分离出两个假定的 HSPC 子集，即 Lin ^- CD45 ^- Sca-1 ^- CD49f ⁺ (CD49f ⁺ ) 和 Lin ^- CD45 ^- Sca-1 ⁺ CD49f ^- (Sca-1 ^{+ ) 细胞。在 CD49f +}中均发现了强大的增殖活性和克隆形成活性和 Sca-1 ⁺细胞通过集落形成测试和细胞周期分析。免疫荧光染色显示CD49f ⁺细胞表达ALB和CK-19，而Sca-1 ⁺细胞仅表达ALB，表明CD49f ⁺细胞具有双潜能，能够分化为肝细胞和胆管细胞。因此，PAS染色显示分化的CD49f ⁺和Sca-1 ⁺细胞合成了糖原，表明它们可以分化成功能性肝细胞。mRNA 表达谱表明 CD49f ⁺和 Sca-1 ⁺细胞显示出与肝祖细胞功能相关的基因的差异表达，例如 Sox9 和 EpCam。此外，两个假定的 HSPC 子集被 DDC 激活，我们发现它们的丰度和增殖随着年龄的增长而增加。总之，我们假设 CD49f ⁺细胞是一种潜在的 HSPC，可用于临床干细胞治疗。