Bone marrow mesenchymal stromal cells stimulate skeletal myoblast proliferation through the paracrine release of VEGF
Year: 2012
Authors: Sassoli C., Pini A., Chellini F., Mazzanti B., Nistri S., Nosi D., Saccardi R., Quercioli F., Zecchi-Orlandini S., Formigli L.
Autors Affiliation: Department of Human Anatomy, Histology and Forensic Medicine, University of Florence, Florence, Italy;
Department of Hematology, Cord Blood Bank, Careggi Hospital, University of Florence, Florence, Italy;
National Institute of Optics (INO), National Research Council (CNR), Sesto Fiorentino, Florence, Italy
Abstract: Mesenchymal stromal cells (MSCs) are the leading cell candidates in the field of regenerative medicine. These cells have also been successfully used to improve skeletal muscle repair/regeneration; however, the mechanisms responsible for their beneficial effects remain to be clarified. On this basis, in the present study, we evaluated in a co-culture system, the ability of bone-marrow MSCs to influence C2C12 myoblast behavior and analyzed the cross-talk between the two cell types at the cellular and molecular level. We found that myoblast proliferation was greatly enhanced in the co-culture as judged by time lapse videomicroscopy, cyclin A expression and EdU incorporation. Moreover, myoblasts immunomagnetically separated from MSCs after co-culture expressed higher mRNA and protein levels of Notch-1, a key determinant of myoblast activation and proliferation, as compared with the single culture. Notch-1 intracellular domain and nuclear localization of Hes-1, a Notch-1 target gene, were also increased in the co-culture. Interestingly, the myoblastic response was mainly dependent on the paracrine release of vascular endothelial growth factor (VEGF) by MSCs. Indeed, the addition of MSC-derived conditioned medium (CM) to C2C12 cells yielded similar results as those observed in the co-culture and increased the phosphorylation and expression levels of VEGFR. The treatment with the selective pharmacological VEGFR inhibitor, KRN633, resulted in a marked attenuation of the receptor activation and concomitantly inhibited the effects of MSC-CM on C2C12 cell growth and Notch-1 signaling. In conclusion, this study provides novel evidence for a role of MSCs in stimulating myoblast cell proliferation and suggests that the functional interaction between the two cell types may be exploited for the development of new and more efficient cell-based skeletal muscle repair strategies.
Journal/Review: PLOS ONE
Volume: 7 (7) Pages from: e37512 to: e37512
More Information: This work was supported by funds from the University of Florence (to LF, SZO, CS) and Italian Ministry of University and Research (MIUR-PRIN 2008 to SZO). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.KeyWords: Animals; Bone Marrow Cells; Cell Line; Cell Proliferation; Coculture Techniques; Mesenchymal Stromal Cells; Mice; Myoblasts, Skeletal; Paracrine Communication; Vascular Endothelial Growth Factor ADOI: 10.1371/journal.pone.0037512Citations: 83data from “WEB OF SCIENCE” (of Thomson Reuters) are update at: 2024-11-17References taken from IsiWeb of Knowledge: (subscribers only)Connecting to view paper tab on IsiWeb: Click hereConnecting to view citations from IsiWeb: Click here