Fig. 4.
Qualitative and quantitative analysis of MSCmix- and NCSCmix-secretome. NCSCmix- and MSCmix-conditioned media (CM) are prepared by culturing MSCmix and NCSCmix in serum-free DMEM for 24 h (with or without stimulation by 1 μg/mL LPS: LPSMSC-CM and LPSNCSC-CM) and analyzed by Mouse Cytokine Array (a). Qualitative analysis of arrays revealed the presence of different chemokines and cytokines, especially present in MSC-secretome (b), and their concentration in each CM sample was assessed by ELISA assays. We observed an increased concentration of G-CSF (c), CXCL1 (d), CCL5 (e), IL-6 (f) in LPSMSC-CM, whereas CXCL12 was present in both MSC-CM and LPSMSC-CM (g). CXCL2 was present in very low concentrations in LPSMSC-CM and LPSNCSC-CM (h). CCL2 was more concentrated in NCSC-CM than in MSC-CM, but increased in both groups when prestimulated with LPS (i). TIMP-1 was equally secreted in all conditions (j), while M-CSF (k) and CXCL10 (l) were present in variable concentrations. (n = 5 to 7, one-way ANOVA and HSD post-test, *p < 0.05, **p < 0.01, ***p < 0.001). For values, see Additional file 1: Table S1. MSC mesenchymal stem cell, NCSC neural crest stem cell, DMEM Dulbecco’s modified Eagle’s medium, LPS lipopolysaccharide, ELISA enzyme-linked immunosorbent assay, G-CSF granulocyte colony-stimulating factor, CXCL C-X-C motif ligand, CCL C-C motif ligand, IL-6 interleukin-6, TIMP1, tissue inhibitor of metalloproteinases 1, ANOVA analysis of variance, HSD honestly significant difference