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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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Related in: MedlinePlus

MSC in a fibrinogen–thrombin matrix were implanted subcutaneously on the sheep’s back (groups 1–3). Explants were harvested after 2 weeks (shown in A and B) and 4 weeks (shown in C and D) to investigate apoptosis and sufficient DiI labelling of the implanted MSC. Constructs with expanded MSC are shown in (A) and (C) versus constructs with directly auto-transplanted MSC shown in (B) and (D). DiI+ cells (red) were identified at all time-points in the subcutaneous implants. The staining intensity of expanded MSC was stronger in comparison to directly auto-transplanted MSC. Apoptotic cells (TUNEL assay) are shown in green. Both expanded and directly auto-transplanted cells had a decreasing apoptosis during the implantation period.
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fig03: MSC in a fibrinogen–thrombin matrix were implanted subcutaneously on the sheep’s back (groups 1–3). Explants were harvested after 2 weeks (shown in A and B) and 4 weeks (shown in C and D) to investigate apoptosis and sufficient DiI labelling of the implanted MSC. Constructs with expanded MSC are shown in (A) and (C) versus constructs with directly auto-transplanted MSC shown in (B) and (D). DiI+ cells (red) were identified at all time-points in the subcutaneous implants. The staining intensity of expanded MSC was stronger in comparison to directly auto-transplanted MSC. Apoptotic cells (TUNEL assay) are shown in green. Both expanded and directly auto-transplanted cells had a decreasing apoptosis during the implantation period.

Mentions: A TUNEL assay was performed to analyse apoptosis. In the first and second week a higher apoptosis could be detected compared to later explantation time-points (Fig. 3A–D, A/B 2 weeks, C/D 4 weeks, A/C expanded MSC, B/D directly auto-transplanted MSC), while the apoptosis decreased from week 1 to week 2. At later time-points only single apoptotic cells were visible. Proliferation was assessed by Ki67 staining. A constant proliferation could be detected over all explantation time-points (Fig. 4A–D, A/B 2 weeks, C/D 4 weeks, A/C expanded MSC, B/D directly auto-transplanted MSC). Despite different marker expression profiles, both cell isolation protocols, directly auto-transplanted or expanded MSC, displayed a constant proliferation and low apoptosis. Therefore, both populations were regarded as suitable for the following implantation studies comparing directly auto-transplanted versus expanded MSC aiming at bone tissue engineering.


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)

MSC in a fibrinogen–thrombin matrix were implanted subcutaneously on the sheep’s back (groups 1–3). Explants were harvested after 2 weeks (shown in A and B) and 4 weeks (shown in C and D) to investigate apoptosis and sufficient DiI labelling of the implanted MSC. Constructs with expanded MSC are shown in (A) and (C) versus constructs with directly auto-transplanted MSC shown in (B) and (D). DiI+ cells (red) were identified at all time-points in the subcutaneous implants. The staining intensity of expanded MSC was stronger in comparison to directly auto-transplanted MSC. Apoptotic cells (TUNEL assay) are shown in green. Both expanded and directly auto-transplanted cells had a decreasing apoptosis during the implantation period.
© Copyright Policy
Related In: Results  -  Collection

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fig03: MSC in a fibrinogen–thrombin matrix were implanted subcutaneously on the sheep’s back (groups 1–3). Explants were harvested after 2 weeks (shown in A and B) and 4 weeks (shown in C and D) to investigate apoptosis and sufficient DiI labelling of the implanted MSC. Constructs with expanded MSC are shown in (A) and (C) versus constructs with directly auto-transplanted MSC shown in (B) and (D). DiI+ cells (red) were identified at all time-points in the subcutaneous implants. The staining intensity of expanded MSC was stronger in comparison to directly auto-transplanted MSC. Apoptotic cells (TUNEL assay) are shown in green. Both expanded and directly auto-transplanted cells had a decreasing apoptosis during the implantation period.
Mentions: A TUNEL assay was performed to analyse apoptosis. In the first and second week a higher apoptosis could be detected compared to later explantation time-points (Fig. 3A–D, A/B 2 weeks, C/D 4 weeks, A/C expanded MSC, B/D directly auto-transplanted MSC), while the apoptosis decreased from week 1 to week 2. At later time-points only single apoptotic cells were visible. Proliferation was assessed by Ki67 staining. A constant proliferation could be detected over all explantation time-points (Fig. 4A–D, A/B 2 weeks, C/D 4 weeks, A/C expanded MSC, B/D directly auto-transplanted MSC). Despite different marker expression profiles, both cell isolation protocols, directly auto-transplanted or expanded MSC, displayed a constant proliferation and low apoptosis. Therefore, both populations were regarded as suitable for the following implantation studies comparing directly auto-transplanted versus expanded MSC aiming at bone tissue engineering.

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