Directly auto-transplanted mesenchymal stem cells induce bone formation in a ceramic bone substitute in an ectopic sheep model.
Bottom Line: Bone matrix proteins were up-regulated in constructs following direct auto-transplantation and in expanded MSC as well as in BMP-2 constructs.Up-regulation was detected using immunohistology methods and RT-PCR.Dense vascularization was demonstrated by CD31 immunohistology staining in all three groups.
Affiliation: Department of Plastic and Hand Surgery, University Hospital of Erlangen, Friedrich-Alexander-University of Erlangen-Nürnberg, Erlangen, Germany.Show MeSH
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Mentions: Sheep MSC were successfully isolated from sheep bone marrow by puncture of the iliac crest. MSC could be isolated using Ficoll gradient centrifugation and plastic adherence selection. In vitro MSC showed a sufficient proliferation. Further characterization by RT-PCR analysis revealed expression of sheep MSC markers CD29, CD44 and CD166 (Fig. 1A). FACS analysis confirmed this observation on the protein level (Fig. 1B–D). Expanded MSC showed a different marker expression profile compared to directly auto-transplanted MSC. Whereas expanded MSC had a strong CD29, CD44 and CD166 expression on protein level, directly auto-transplanted MSC seemed to have only a strong CD44 expression and a weak CD29 and CD166 expression. Differences in expression of CD29, CD44 and CD166 in directly auto-transplanted MSC compared to expanded MSC were less evident on mRNA level. As indicated by increased CD45 expression, ratio of hematopoietic cells was higher in directly auto-transplanted MSC as compared to expanded MSC.
Affiliation: Department of Plastic and Hand Surgery, University Hospital of Erlangen, Friedrich-Alexander-University of Erlangen-Nürnberg, Erlangen, Germany.