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Fig. 4 | Stem Cell Research & Therapy

Fig. 4

From: Placental chorionic plate-derived mesenchymal stem cells ameliorate severe acute pancreatitis by regulating macrophage polarization via secreting TSG-6

Fig. 4

GdCl3 and MCLs selectively induced apoptosis of M1 or M2 macrophages. To investigate the effects of GdCL3 and MCLs on macrophages with different polarization phenotypes in vitro, we isolated and cultured macrophages from the bone marrow and induced the macrophages into M1 and M2 phenotypes. A Flow cytometry analysis of the apoptosis of macrophages with different polarization phenotypes after 48-h intervention with gadolinium (III) chloride (GdCl3) or mannosylated clodronate-encapsulated liposomes (MCLs). B, C Quantitative results of the apoptosis rate when macrophages with different polarization phenotypes were intervened by GdCl3 or MCLs (n = 3). The total apoptotic number of macrophages is from AnV+ and Anv/PI+. To confirm that we successfully induced bone marrow cells into M1 and M2 macrophages in vitro, the polarization phenotype of macrophages was analyzed by flow cytometry (D) and RT-qPCR (G, H), CD163+ macrophages are considered as M2 macrophages, and CD86+ macrophages are considered as M1 macrophages. E, F Flow cytometric analysis of macrophages with different polarization phenotypes (n = 3). G The mRNA expression levels of M1 marker genes (IL1-β, TNF-α, and iNOS) in macrophages (n = 3). H The mRNA expression levels of M2 marker genes (Arg-1, IL-10, and CD163) in macrophages (n = 3). To observe the effect of CP-MSCs on macrophage polarization in vivo, all rats were sacrificed 72h after the first CP-MSC transplantation, liver and pancreas tissues were collected, and then the polarization phenotype of macrophages was detected by immunofluorescence staining. Representative immunofluorescence staining of the polarized phenotype of macrophages in the pancreas (I) and liver tissues (J). Scale bars, 50 μm. K Quantification of M2 macrophages (CD163+) and M1 macrophages (iNOS+) in the pancreas and liver tissue, the numbers of immunopositive cells represent the mean ± SEM of at least 5 fields (n = 5). All graphs show mean ± SEM. Significance is indicated as follows: *p < 0.05; **p < 0.01; ***p < 0.001; NS, no significant. M0 represents unpolarization-induced bone marrow macrophages; M1 represents M1 polarization-induced bone marrow macrophages; M2 represents M2 polarization-induced bone marrow macrophages

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