Therapeutic Benefits of Induced Pluripotent Stem Cells in Monocrotaline-Induced Pulmonary Arterial Hypertension
Fig 6
iPSC-based treatment suppressed inflammation in polarized classical human macrophages.
The effects of murine iPSCs on inflammation were evaluated in the context of co-culture with polarized classical human macrophages. Primitive human macrophages were differentiated from PBMCs and subsequently polarized (via the stimulation of LPS and IFNγ) to become proinflammatory M1 macrophages. To examine paracrine effects of iPSCs in vitro, cells were separated using a permeable membrane (A), thereby mimicking the conditioned media treatment in rats. (B) Secretion of the inflammatory cytokines TNFα and IL-1β was measured via ELISA. Data were accumulated from 3–5 independent assays. (C) The immunocytochemical analysis of inflammatory markers CD68 and MHC-II was performed on co-cultured iPSCs and M1 macrophages. CD68 and MHC-II (Texas Red) and iPSC-specific marker SSEA-1 (FITC) are shown. Nuclei were counterstained with Hoechst 33342. (D) Genes specifically expressed in M1 macrophages were compared across groups of iPSC-based treatments using semi-quantitative RT-PCR. The results are represented as the means ± SD; values were normalized according to 36b4 mRNA levels and represented as the fold change relative to the PBS control group. *p<0.05 vs. PBS group; †p<0.05 vs. MCT group. Scale bar = 10 μm.