Open Access Research article

Characterization of in vivo chemoresistant human hepatocellular carcinoma cells with transendothelial differentiation capacities

Christian Marfels, Miriam Hoehn, Ernst Wagner and Michael Günther*

Author Affiliations

Pharmaceutical Biotechnology, Department of Pharmacy, Ludwig-Maximilians-Universität, Butenandtstrasse 5-13, D-81377, Munich, Germany

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BMC Cancer 2013, 13:176  doi:10.1186/1471-2407-13-176

Published: 2 April 2013

Additional files

Additional file 1: Table S1:

Abbreviations of cell lines and their origin.

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Additional file 2: Table S2:

Used antibodies for immunohistochemistry.

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Additional file 3: Table S3:

Used pairs of primer for qRT-PCR.

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Additional file 4: Figure S1:

In vitro control of new generated cell lines. HUH-wt (A), HUH-PAS (B) and HUH-REISO (C) showed no differences in morphology by transmitted light microscopy. Moreover, in FACS analysis cells showed human origin as all cells of HUH-wt (D), HUH-PAS (E) and HUH-REISO (F) were positive in the staining for human epidermal growth factor receptor (hEGF-R), indicated by the shifted red curve in comparison to the correspondent control antibody (white curve).

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Additional file 5: Figure S3:

Hoechst-blood-flow comparison of HUH-REISO vs HUH-PAS after therapy. Intratumoral functional blood flow was visualized by intravenous application of Hoechst 33258 dye (blue). Cryosections (5μm) of HUH-PAS control tumors after 2×CPA treatment (A-C) showed a clear diminished functional blood flow in comparison to treated HUH-REISO tumors (D-F). To ensure to have tumor-material for analysis, the beginning of treatment was late for HUH-PAS tumors, when tumors reached an average volume of 254 mm3.

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Additional file 6: Figure S2:

Software based analysis of Matrigel assay. The software based analysis system projected a mask on pictures from transmitted light microscopy of matrigel assay (same pictures as described in Figure 9D, E and F). In white are confluent areas, in blue are nodular structures coloured and built counted tubes with orange and red coloured areas. The tube skeleton is indicated in the thin white lines and their crossing points are defined as branching points. As HUH-wt (A) and HUH-PAS (B) revealed no tube formation, the projected mask is hardly coloured (D and E). HUH-REISO (C) showed low amount of confluent white area and accordingly contained all other aspects of functional tube formation in the mask (F).

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