CTGF directs fibroblast differentiation from human mesenchymal stem/stromal cells and defines connective tissue healing in a rodent injury model
Chang H. Lee, … , Eduardo K. Moioli, Jeremy J. Mao
Chang H. Lee, … , Eduardo K. Moioli, Jeremy J. Mao
Published August 2, 2010
Citation Information: J Clin Invest. 2010;120(9):3340-3349. https://doi.org/10.1172/JCI43230.
View: Text | PDF | Erratum
Research Article

CTGF directs fibroblast differentiation from human mesenchymal stem/stromal cells and defines connective tissue healing in a rodent injury model

Abstract

Fibroblasts are ubiquitous cells that demonstrate remarkable diversity. However, their origin and pathways of differentiation remain poorly defined. Here, we show that connective tissue growth factor (CTGF; also known as CCN2) is sufficient to induce human bone marrow mesenchymal stem/stromal cells (MSCs) to differentiate into fibroblasts. CTGF-stimulated MSCs lost their surface mesenchymal epitopes, expressed broad fibroblastic hallmarks, and increasingly synthesized collagen type I and tenacin-C. After fibroblastic commitment, the ability of MSCs to differentiate into nonfibroblastic lineages — including osteoblasts, chondrocytes, and adipocytes — was diminished. To address inherent heterogeneity in MSC culture, we established 18 single MSC–derived clones by limiting dilution. CTGF-treated MSCs were α-SMA–, differentiating into α-SMA+ myofibroblasts only when stimulated subsequently with TGF-β1, suggestive of stepwise processes of fibroblast commitment, fibrogenesis, and pathological fibrosis. In rats, in vivo microencapsulated delivery of CTGF prompted postnatal connective tissue to undergo fibrogenesis rather than ectopic mineralization. The knowledge that fibroblasts have a mesenchymal origin may enrich our understanding of organ fibrosis, cancer stroma, ectopic mineralization, scarring, and regeneration.

Authors

Chang H. Lee, Bhranti Shah, Eduardo K. Moioli, Jeremy J. Mao

×

Figure 4

Myofibroblastic differentiation of MSC-derived fibroblastic cells by TGF-β1.

Options: View larger image (or click on image) Download as PowerPoint
Myofibroblastic differentiation of MSC-derived fibroblastic cells by TGF...
(A–D) Native MSCs (A) or MSC-derived fibroblasts by CTGF treatment (C) expressed little α-SMA. Upon TGF-β1 treatment, native MSCs still expressed little α-SMA (B), but MSC-derived fibroblasts readily expressed α-SMA+ microfilaments (D). (E–H) Flow cytometry confirmed the virtual absence of α-SMA expression in MSCs (E) or MSC-derived fibroblasts (G). In contrast, 31.9% of MSC-derived fibroblasts (H), but only 1.8% of native MSCs (F), gained α-SMA phenotype after TGF-β1 stimulation. (I–L) Collagen gel contraction assay showed that MSCs with sequential administration of CTGF (4 weeks) and TGF-β1 (1 week) yielded the most significant contraction (I), compared with moderate contraction upon CTGF stimulation alone (J) or TGF-β1 stimulation alone of native MSCs (K). MSCs without either CTGF or TGF-β1 stimulation yielded the least contraction (L). (M) Quantitatively, sequential stimulation of MSCs by CTGF and TGF-β1 yielded the most significant collagen gel contraction (P < 0.05). Scale bars: 100 μm. Data represent mean ± SD.