Figure 4. MB231-LEC recruit pericytes in vivo.
High-concentrate matrigel containing LEC or HUVEC (2 × 106/gel) and heparin (10 units/gel) was injected subcutaneously. TCM or SFM (50 μl/injection) was subcutaneously dosed daily for 10 days, the mice were euthanized, and the gel plugs were excised and analyzed. (a) Gel plugs were sectioned and probed with anti-alpha-SMA (αSMA) antibodies. αSMA positive cells per 12 randomly selected areas were quantified by using ImageJ. The LEC-TCM group shows significant infiltration of the αSMA positive cells, compared to the LEC-SFM group (*P = 0.031). (b) Representative confocal images for αSMA-positive area in four different groups of plugs (HUVEC-TCM, LEC-TCM, LEC-SFM, No cell-TCM). Scale bars in the whole images and in the enlarged images represent 5,000 μm and 1,000 μm, respectively. (c) To confirm the pericyte infiltration and blood vessel coverages by the pericytes, four different plug samples were co-stained with anti-desmin and anti-mCD31 antibodies. LEC-included matrigel plugs in animals treated with TCM showed higher coverage of the blood vessels by pericytes, compared to TCM-treated HUVEC, SFM-treated LEC, and TCM-treated no cell groups. Anti-hVEGFR3 antibodies (blue) were used to detect human LEC that are included in the gel plugs. Only LEC-TCM and LEC-SFM group showed LEC. Scale bars represent 200 μm. Data (a) are reported as mean ± s.e.m.