Skip to main content
Figure 3 | Stem Cell Research & Therapy

Figure 3

From: Hypoxia-preconditioned mesenchymal stem cells attenuate bleomycin-induced pulmonary fibrosis

Figure 3

Hypoxia-preconditioned mesenchymal stem cells attenuate extracellular matrix production through transforming growth factor-β1-mediated Akt signaling. In the assessment of the anti-fibrotic effects of mesenchymal stem cells (MSCs), transforming growth factor (TGF)-β1-stimulated MRC-5 fibroblasts were used. (A) Co-culture experiments were performed in which mock control (Ctrl, culture medium only), TGF-β1 control, normoxia-preconditioned MSCs (NP-MSCs) or hypoxia-preconditioned MSCs (HP-MSCs) were seeded in the upper chamber, and TGF-β1 (1 ng/ml)-treated MRC-5 fibroblast cells were seeded in the lower chamber. After 24 hours incubation, quantitative real-time RT-PCR was performed to examine the fibronectin mRNA levels in MRC-5 cells. (B) Western blot analysis of fibronectin and phosphorylated Akt in TGF-β1-treated MRC-5 cells after incubation with MSC-conditioned media (CM) for 24 hours. (C) Co-culture experiments were performed in which mock Ctrl, TGF-β1 Ctrl, HP-Ctrl (hypoxia-pretreated MSCs transfected with control scramble short hairpin RNA (shRNA) plasmid) or HP-shHGF were seeded in the upper chamber, and TGF-β1 (1 ng/ml)-treated MRC-5 fibroblast cells were seeded in the lower chamber. After 24 hours incubation, quantitative real-time RT-PCR was performed to examine the fibronectin mRNA levels in MRC-5 cells. (D) MRC-5 cells were preincubated with or without PHA-665752 (1 μM) for 24 hours, and then cells were treated with or without HP-MSC CM for 24 hours. Quantitative real-time RT-PCR was performed to examine changes in the fibronectin mRNA levels in the MRC-5 cells. Values were normalized to the β-actin gene and are expressed relative to the control (Ctrl) group. **P < 0.01, ***P < 0.001. HGF, hepatocyte growth factor.

Back to article page