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Version of Record online: 22 OCT 2012
Copyright © 2012 AlphaMed Press
Volume 30, Issue 11, pages 2460–2471, November 2012
How to Cite
van Gastel, N., Torrekens, S., Roberts, S. J., Moermans, K., Schrooten, J., Carmeliet, P., Luttun, A., Luyten, F. P. and Carmeliet, G. (2012), Engineering Vascularized Bone: Osteogenic and Proangiogenic Potential of Murine Periosteal Cells. STEM CELLS, 30: 2460–2471. doi: 10.1002/stem.1210
Author contributions: N.v.G.: conception and design, collection and assembly of data, data analysis and interpretation, and manuscript writing; S.T., S.J.R., and K.M.: collection and assembly of data; J.S.: conception and design; P.C. and A.L.: provision of study material; F.P.L.: conception and design, data analysis and interpretation, and provision of study material; G.C.: conception and design, data analysis and interpretation, manuscript writing, and final approval of manuscript.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS August 22, 2012.
- Issue online: 22 OCT 2012
- Version of Record online: 22 OCT 2012
- Accepted manuscript online: 21 AUG 2012 08:32AM EST
- Manuscript Accepted: 25 JUL 2012
- Manuscript Received: 4 JUN 2012
- Fund for Scientific Research Flanders. Grant Numbers: FWO; G.0500.08, G.0982.11
- Flemish Government
- Long-term Structural Funding—Methusalem
- European Commission. Grant Number: FP7-StG-IMAGINED 203291
- KU Leuven (IOF Knowledge Platform “Prometheus”. Grant Number: IOFKP/07/004
- Stem Cell Institute of Leuven
- Agency for Innovation by Science and Technology in Flanders (IWT)
- Prometheus, the Leuven Research & Development Division of Skeletal Tissue Engineering of the KU Leuven
Additional Supporting Information may be found in the online version of this article.
|sc-12-0509_sm_SupplFigure1.pdf||882K||Figure S1. Isolation of mPDC: embedding of epiphyses and enzymatic digest. (A-C): H&Estained sections showing murine tibias before digest (A), after enzymatic digest (B) and after embedding of the epiphyses in low melting point agarose and enzymatic digest (C). Black arrows indicate remaining tissue from the joint and tendon present on the epiphysis. (A1-C1): Magnification of the boxed areas in A-C showing release of perichondrial cells after enzymatic digest, which was precluded when epiphyses were first embedded in agarose (black arrow). (A2-C2): H&E-stained sections showing the periosteal surface of the diaphysis (black arrow) of the tibia before digest (A2) or after enzymatic digest without (B2) or with (C2) epiphyses embedded in low melting point agarose. Scale bars = 500μm in A, B, C; 200μm in A1, B1, C1, A2, B2, C2. Abbreviations: H&E: hematoxylin and eosin.|
|sc-12-0509_sm_SupplFigure2.pdf||337K||Figure S2. mPDC stabilize networks formed by endothelial cells in vitro. (A, B): CMFDAlabeled mPDC were co-cultured with CM-DiI-labeled HUVEC or hBOEC on MatrigelTM for 24h. Brightfield (A) and fluorescent (B) microscopic analysis of cultures of endothelial cells alone (left), together with mPDC (middle) or of mPDC alone (right) showing formation of robust pseudovascular networks in the co-cultures, with mPDC localized predominantly at the outside of the structures, around the endothelial cells. Scale bar = 200μm. Abbreviations: hBOEC: human blood outgrowth endothelial cells; HUVEC: human umbilical vein endothelial cells; mPDC: murine periosteum-derived cells.|
|sc-12-0509_sm_SupplFigure3.pdf||366K||Figure S3. Co-implantation of mPDC and HUVEC results in the formation of mature, perfused blood vessels. (A-B): In vivo co-implantation of mPDC and HUVEC in a 60:40 ratio in collagen gels for 7 days. H&E-stained sections (A) showing the presence of small (A1, A1′) and large (A2, A2′) blood vessels filled with red blood cells, confirming their connection to the host vasculature. Immunohistochemical staining of HUVEC with UEA-lectin in combination with green autofluorescence of blood erythrocytes (B) indicates that the blood vessels formed by the implanted HUVEC are functional. The section was counterstained with Hoechst to visualize cell nuclei. Scale bars = 500μm in A; 100μm in A1; 50μm in A1′, A2, B; 20μm in A2′. Abbreviations: H&E: hematoxylin and eosin, HUVEC: human umbilical vein endothelial cells; mPDC: murine periosteum-derived cells; UEA-lectin: Ulex europaeus agglutinin-lectin.|
|sc-12-0509_sm_SupplTable1.pdf||77K||Supplementary Table 1|
|sc-12-0509_sm_SupplTable2.pdf||65K||Supplementary Table 2|
|sc-12-0509_sm_SupplTable3.pdf||31K||Supplementary Table 3|
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