Fibronectin stimulates the osteogenic differentiation of murine embryonic stem cells

Abstract

Conditioned medium from human hepatocarcinoma cells (HepG2-CM) has been shown to stimulate the osteogenic/chondrogenic differentiation of murine embryonic stem cells (mESCs). HepG2-CM is considered to contain visceral endoderm (VE)-like signals and attempts have recently been made to characterize it, using proteomic profiling, with fibronectin being identified as one promising candidate. Herein, we investigated whether fibronectin is able to mimic the activities of HepG2-CM during the osteogenic differentiation of mESCs. Specifically, the addition of RGD peptides and heparin in HepG2-CM significantly reduced the growth- and adhesion-promoting effects of HepG2-CM, in addition to suppressing its osteogenic-inductive activity. Furthermore, direct addition of fibronectin to basal medium was able to reproduce, at least partially, the function of HepG2-CM. In particular, fibronectin induced the early onset of osteogenic differentiation in mESCs, as confirmed by gene expression of osteogenic markers, and resulted in the three-fold higher calcium deposition at day 11 of osteogenic culture compared to the control group. These data clearly suggest that fibronectin contributes to the biological activities of HepG2-CM and plays a stimulatory role during the process of osteogenesis in mESCs. Copyright © 2015 John Wiley & Sons, Ltd.

Collagen Promotes Higher Adhesion, Survival and Proliferation of Mesenchymal Stem Cells

“…These results suggest that COL is a suitable cell expansion matrix to retain high osteogenic differentiation potential of MSC and provide a synergistic effect in promoting osteogenic differentiation in the presence of induction factors prior to in vivo administration. Similar increase in osteogenic differentiation on COL surface was also reported by Linsley et al and others [ 21 , 22 , 24 – 26 , 28 , 47 ]. Although chondrogenic differentiation is an important property of MSC, it was not tested in the current study as chondrogenic differentiation in vitro is usually performed as a pellet culture where there was no scope for cell to cell-culture-surface contact.…”

“…Although the matrix rigidity and elasticity were not tested in our study, the results show that differential mechano-sensing and signaling was promoted in cells cultured on different matrices. Even though other studies have shown the usefulness of COL and FBN as suitable extracellular matrix for osteogenic differentiation of MSC [ 21 , 22 , 24 – 26 , 28 , 47 ], our study specifies their suitability in enhancing cell attachment, migration, cell survival and maintaining their self-renewal and differentiation potential. Taken together, our results suggest that COL might be employed as a matrix during in vitro cell expansion to maintain their osteogenic differentiation potential and coating in vivo scaffolds for increased cell adhesion, proliferation, survival during stress conditions and migration.…”

RGD‐functionalized polyurethane scaffolds promote umbilical cord blood mesenchymal stem cell expansion and osteogenic differentiation

“…Efficient bone tissue engineering strategies require the combination of bioactive materials carrying extracellular matrix (ECM) cues with stem cells (Reddi, ). ECM has been used to increase the bioactivity of polymers, such as PU, and to produce biomaterials capable of regulating stem cell proliferation and differentiation via signals such as cell adhesion (Ahmed & Ffrench‐Constant, ; Kang et al, ). ECM peptides can be synthesized on demand under Good Manufacturing Practice (GMP) conditions eliminating infection risks and avoiding potential immune reactions to animal‐derived proteins such as collagen (Hersel, Dahmen, & Kessler, ).…”

“…ECM peptides can be synthesized on demand under Good Manufacturing Practice (GMP) conditions eliminating infection risks and avoiding potential immune reactions to animal‐derived proteins such as collagen (Hersel, Dahmen, & Kessler, ). The arginine‐glycine‐aspartic acid (RGD) sequence was initially identified as the adhesion motif of fibronectin but is now known to be present in a wide variety of ECM proteins essential for cell adhesion and differentiation (Kang et al, ; Ruoslahti, ). Biomaterial surface functionalization with peptides, such as RGD, is advantageous compared with ECM proteins due to the fact that bioactive sites of a whole protein may not be available for integrin binding (Pierschbacher & Ruoslahti, ).…”

Therapeutic effects of Salvia miltiorrhiza injection combined with telmisartan in patients with diabetic nephropathy by influencing collagen IV and fibronectin: A case-control study

“…Similar to integrin, FN can bind ECM components such as collagen, fibrin, and heparan sulfate proteoglycans ( 13 ). Furthermore, FN plays a major role in cell adhesion, growth, migration, and differentiation, and it is important for wound healing and embryonic development ( 14 – 16 ). Altered FN expression, degradation, and/or organization have been associated with a number of pathological changes, including fibrosis and cancer ( 17 , 18 ).…”