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 Article has an altmetric score of 7

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 2

CTGF-derived fibroblasts are a stable population.

Options: View larger image (or click on image) Download as PowerPoint
CTGF-derived fibroblasts are a stable population. (A–F) MSC-derived fibr...
(A–F) MSC-derived fibroblasts (MSC-Fb) by CTGF treatment for 4 weeks showed minimal capacity to further differentiate into osteoblasts (A), chondrocytes (B), or adipocytes (C). In contrast, native MSCs, without CTGF treatment, readily differentiated into osteoblasts (D), chondrocytes (E), and adipocytes (F). (G–I) von Kossa staining was negative in CTGF-treated MSCs (H), just as MSCs without CTGF treatment (G). (I) In contrast, MSCs subjected to osteogenic stimulation readily differentiated into osteogenic cells that elaborated minerals. (J–L) Safranin O staining was negative in CTGF-treated MSCs (K), just as in MSCs without CTGF treatment (J). (L) In contrast, MSCs subjected to chondrogenic stimulation readily differentiated into chondrogenic cells that were safranin O positive. (M) Quantitatively, MSCs under osteogenic stimulation (MSC-Ob) elaborated significantly more calcium than did MSCs with or without CTGF treatment (n = 5). (N) In parallel, MSCs under chondrogenic stimulation (MSC-Ch) produced significantly more glycosaminoglycans (GAG) than MSCs with or without CTGF treatment (n = 5). Scale bars: 100 μm (A, B, D, E, and G–L); 50 μm (C and F). Data represent mean ± SD. *P < 0.05; **P < 0.01.