IGF-I and IGF-II stimulate directed cell migration of bone-marrow-derived human mesenchymal progenitor cells

doi:10.1016/j.bbrc.2006.05.034

Biochemical and Biophysical Research Communications

Volume 345, Issue 3, 7 July 2006, Pages 1177-1183

https://doi.org/10.1016/j.bbrc.2006.05.034 Get rights and content

Abstract

Insulin-like growth factors (IGFs) are known to be key regulators of bone growth, remodeling, and repair. Since all these processes depend on the recruitment of cells with the potential to be committed to the osteoblastic lineage, we studied possible effects of IGF-I and -II on migration of human mesenchymal progenitor cells (MPC) using a modified Boyden chamber assay. The results were compared to those of primary osteoblasts and in vitro-osteogenic-differentiated MPC. IGF-I and -II stimulated cell migration of all these cell populations in a dose-dependent manner from 1 to 100 ng/mL. The maximal chemotactic index (CI) was 4–5 for MPC and primary osteoblasts and about 3 for in vitro-differentiated MPC. Checkerboard analysis revealed that IGFs stimulated true directed cell migration (chemotaxis) and not simply chemokinesis. Addition of an antibody against the type I IGF receptor (αIR3) completely abolished (MPC) or markedly reduced (primary osteoblasts) the chemotactic effects of each of the IGFs. IGFBP-3 itself had no direct effect, while IGFBP-5 stimulated MPC migration at concentrations of 80 and 160 ng/mL. Parallel application of IGFBP-3 had borderline inhibitory effects while the addition of 40 ng/mL of IGFBP-5 enhanced the chemotactic effect of IGF-I on MPC. In conclusion, our results show that IGF-I and -II are chemotactic factors for MPC and indicate that IGFBP-5 both modulates the IGF-I effect and directly stimulates migration of human mesenchymal progenitor cells.

Section snippets

Materials and methods

Cell culture. With informed consent, following the guidelines from the Ethics Committee of the University of Ulm, bone marrow aspirates from six donors (aged 20 to 30 years) were plated and cultured in Dulbecco’s minimal essential medium (DMEM; Biochrom, Germany) containing 10% FCS as described previously [5]. Experiments were performed only in the first four cell passages. In previous studies, we showed that this technique of cell isolation maintains the multipotency phenotype, deduced from

Results

Human MPC used for the chemotaxis assays did not express markers of differentiated osteoblasts (osteocalcin), chondrocytes (collagen type II) or adipocytes (leptin). Supplementation with dexamethasone, ascorbic acid, and β-glycerophosphate led to osteogenic differentiation within 21 days, indicated by the expression of lineage-typical gene expression profiles (Fig. 1).

IGF-I induced a dose-dependent migratory response in human bone marrow derived MPC (Fig. 2), with the maximal value of the CI

Discussion

A chemotactic effect of IGF-I and -II has been previously described for primary human osteoblasts [12]. We have confirmed these results and extended the knowledge on the regulation of cell migration in progenitor cells of the osteoblastic lineage. This is of special importance since in recent years it has turned out that locally available, bone-marrow-derived or even circulating, mesenchymal or osteo-progenitor cells are crucially involved in processes such as bone growth, remodeling or repair

Acknowledgments

The authors thank Eli Lilly (Greenfield, Indiana, USA) for providing human IGF-I and IGF-II and Mr. Giovanni Ravalli for expert technical assistance.

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^☆: This work was funded in part by the “Land Baden-Württemberg.”

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IGF-I and IGF-II stimulate directed cell migration of bone-marrow-derived human mesenchymal progenitor cells☆

Abstract

Section snippets

Materials and methods

Results

Discussion

Acknowledgments

J. Orthop. Res.

Bone

Bone

Metabolism

Biochem. Biophys. Res. Commun.

Exp. Mol. Pathol.

Mol. Cell. Endocrinol.

Growth Horm. IGF Res.

Bone

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

J. Biol. Chem.

J. Invest. Dermatol.

Mesenchymal stem cells reside within the connective tissues of many organs

Dev. Dyn.

Mesenchymal stem cells

J. Orthop. Res.

BMP-2, BMP-4, and PDGF-bb stimulate chemotactic migration of primary human mesenchymal progenitor cells

J. Cell. Biochem.

To go or not to go: migration of human mesenchymal progenitor cells stimulated by isoforms of PDGF

J. Cell. Biochem.

VEGF-A and PlGF-1 stimulate chemotactic migration of human mesenchymal progenitor cells

Biochem. Biophys. Res. Commun.

Normal growth and development in the absence of hepatic insulin-like growth factor I

Proc. Natl. Acad. Sci. USA