Lymphatic Drainage-Promoting Effects by Engraftment of Artificial Lymphatic Vascular Tissue Based on Human Adipose Tissue-Derived Mesenchymal Stromal Cells in Mice

Figure 2

The effects of hASCs on artificial lymphatic vascular network formation. (a, b) The lymphatic vascular formation-promoting effect of hASCs was evaluated by addition to upper or lower layer of FbLT (green: HDLECs; grey: fibroblasts; orange: hASCs). The lymphatic vascular structure was immunostained for CD31, and then the density of the vascular structure along Z axis was quantified as shown in the graphs (b). Green dotted lines indicate the main peak of vascular structure upon HDLEC-seeded level. Arrows indicate the extra peaks of the vascular structure. (c) The lymphatic vascular networks of FbLT with and without an addition of 25% hASCs in the upper layer. The networks were immunostained for CD31, and then the confocal image was analyzed using Fiji software; the Z axis level of network structures was colored according to the left color bar. Arrow heads indicate the sprouting of lymphatic vessels in the upper layer. Addition of 25% hASCs resulted in an increase of the sprouting. (d) Quantitative analysis of increased lymphatic vascular structures by addition of hASCs at several amounts (n = 3). ANOVA with Bonferroni post hoc test showed significant difference of lymphatic vascular density by addition of hASCs to the lower layer (0%, 25%, and 100%,  < 0.05). By addition of hASCs to the top layer (0%, 10%, 25%, and 100%), tendency of difference was shown, although it was not significant (NS).

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