Improved method increases sensitivity for circulating hepatocellular carcinoma cells

Figure 1 Flow cytometric analysis. Recovery of spiked HepG2 human liver cancer cells from blood samples using depletion of CD45⁺ leukocytes (white bars) and asialoglycoprotein receptor-positive selection (black bars).

Figure 2 Immunofluorescence with hepatic antibody cocktail. A: Immunofluorescence staining of human hepatocellular carcinoma tissue with anti-asialoglycoprotein receptor (ASPGR; green) and anti-carbamoyl phosphate synthetase 1 (CPS1; red) antibodies and costained with DAPI (blue) (magnification × 400); B: ASGPR⁺ and CPS1⁺ cells were analyzed by flow cytometry of Hep3B human liver cancer cells.

Figure 3 Improved circulating tumor cell detection sensitivity. A: Immunofluorescence staining of circulating tumor cells (CTCs) (arrow) detected in blood from hepatocellular carcinoma patients with antibodies against asialoglycoprotein receptor (ASPGR) and/or carbamoyl phosphate synthetase 1 (CSP1) (red), and CD45 (green) with nuclear DAPI staining (blue) (magnification × 400); B: CTC numbers detected in the same HCC patients using the currently described method of CD45⁺ depletion and ASPGR⁺/CSP1⁺ immunostaining (green squares) and a previously described method with ASPGR⁺ selection followed by staining with cytokeratin and CSP1 antibodies (purple diamonds); C: The current method showed higher detection sensitivity in all patients with > 40 CTCs.