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Directly auto-transplanted mesenchymal stem cells induce bone formation in a ceramic bone substitute in an ectopic sheep model.

Boos AM, Loew JS, Deschler G, Arkudas A, Bleiziffer O, Gulle H, Dragu A, Kneser U, Horch RE, Beier JP - J. Cell. Mol. Med. (2010)

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.

View Article: PubMed Central - PubMed

Affiliation: Department of Plastic and Hand Surgery, University Hospital of Erlangen, Friedrich-Alexander-University of Erlangen-Nürnberg, Erlangen, Germany.

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Sheep MSC were characterized using FACS and RT-PCR analysis. (A) With RT-PCR analysis CD29, CD44 and CD166 expression of MSC could be proofed 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. (B–D) FACS analysis revealed sheep MSC to express CD29, CD44 and CD166. Expanded MSC (B) were negative for the hematopoietic markers CD31 and CD45. Directly auto-transplanted cells (C) had a different expression pattern than expanded MSC. The directly auto-transplanted MSC had a weaker CD29 and CD166 but a stronger CD45 expression. Mean fluorescent indices are shown in (D).
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fig01: Sheep MSC were characterized using FACS and RT-PCR analysis. (A) With RT-PCR analysis CD29, CD44 and CD166 expression of MSC could be proofed 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. (B–D) FACS analysis revealed sheep MSC to express CD29, CD44 and CD166. Expanded MSC (B) were negative for the hematopoietic markers CD31 and CD45. Directly auto-transplanted cells (C) had a different expression pattern than expanded MSC. The directly auto-transplanted MSC had a weaker CD29 and CD166 but a stronger CD45 expression. Mean fluorescent indices are shown in (D).

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.


Directly auto-transplanted mesenchymal stem cells induce bone formation in a ceramic bone substitute in an ectopic sheep model.

Boos AM, Loew JS, Deschler G, Arkudas A, Bleiziffer O, Gulle H, Dragu A, Kneser U, Horch RE, Beier JP - J. Cell. Mol. Med. (2010)

Sheep MSC were characterized using FACS and RT-PCR analysis. (A) With RT-PCR analysis CD29, CD44 and CD166 expression of MSC could be proofed 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. (B–D) FACS analysis revealed sheep MSC to express CD29, CD44 and CD166. Expanded MSC (B) were negative for the hematopoietic markers CD31 and CD45. Directly auto-transplanted cells (C) had a different expression pattern than expanded MSC. The directly auto-transplanted MSC had a weaker CD29 and CD166 but a stronger CD45 expression. Mean fluorescent indices are shown in (D).
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Related In: Results  -  Collection

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fig01: Sheep MSC were characterized using FACS and RT-PCR analysis. (A) With RT-PCR analysis CD29, CD44 and CD166 expression of MSC could be proofed 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. (B–D) FACS analysis revealed sheep MSC to express CD29, CD44 and CD166. Expanded MSC (B) were negative for the hematopoietic markers CD31 and CD45. Directly auto-transplanted cells (C) had a different expression pattern than expanded MSC. The directly auto-transplanted MSC had a weaker CD29 and CD166 but a stronger CD45 expression. Mean fluorescent indices are shown in (D).
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.

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.

View Article: PubMed Central - PubMed

Affiliation: Department of Plastic and Hand Surgery, University Hospital of Erlangen, Friedrich-Alexander-University of Erlangen-Nürnberg, Erlangen, Germany.

Show MeSH
Related in: MedlinePlus