Journal: Journal of Cellular and Molecular Medicine

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

doi: 10.1111/j.1582-4934.2010.01131.x

Figure Lengend Snippet: 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).

Article Snippet: CD31 staining: After antigen retrieval with pH 6 solution (Target Retrieval Solution; Dako Cytomation) in a pressure cooker for 10 min. (Pascal; Dako Cytomation) peroxidase block (CSAII-System; Dako Cytomation) was applied for 15 min., followed by incubation with 10% goat serum (PromoCell GmbH) in PBS (PBS-Dulbecco 1×, Biochrom AG) for 30 min. and protein block with the CSA II-System for 30 min. Then sections were incubated with the primary monoclonal mouse anti-ovine antibody CD31 (Anti-CD31/PECAM-1, MorphoSys UK Ltd., Kidlington, Oxford, UK) at 1:100 diluted in 10% goat serum in PBS for 1 hr.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Journal of Cellular and Molecular Medicine

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

doi: 10.1111/j.1582-4934.2010.01131.x

Figure Lengend Snippet: For determination of the cell type which is qualified best for bone tissue engineering purposes, different groups (expanded versus directly auto-transplanted MSC, groups 8–10) were investigated. In both groups cells were DiI labelled prior to implantation and implanted subcutaneously with or without BMP-2. (A–C) Expanded MSC (A), directly auto-transplanted MSC (B), BMP-2 in combination with directly auto-transplanted MSC (C). DiI-labelled MSC (red) could be found close to β-TCP/HA granules contributing to the newly formed bone parts. In the explants with directly auto-transplanted MSC a higher section of the DiI-labelled cells were found in the connective tissue parts of the constructs compared to the explants with expanded MSC or directly auto-transplanted MSC with BMP-2. (D–F) Sections of constructs of the groups with expanded MSC (D), directly auto-transplanted MSC (E), BMP-2 in combination with directly auto-transplanted MSC (F) were evaluated for vascularization. The constructs in all three groups are well vascularized as shown by CD31 immunohistochemistry (green). Nuclei are counterstained with DAPI (blue).

Article Snippet: CD31 staining: After antigen retrieval with pH 6 solution (Target Retrieval Solution; Dako Cytomation) in a pressure cooker for 10 min. (Pascal; Dako Cytomation) peroxidase block (CSAII-System; Dako Cytomation) was applied for 15 min., followed by incubation with 10% goat serum (PromoCell GmbH) in PBS (PBS-Dulbecco 1×, Biochrom AG) for 30 min. and protein block with the CSA II-System for 30 min. Then sections were incubated with the primary monoclonal mouse anti-ovine antibody CD31 (Anti-CD31/PECAM-1, MorphoSys UK Ltd., Kidlington, Oxford, UK) at 1:100 diluted in 10% goat serum in PBS for 1 hr.

Techniques: Construct, Immunohistochemistry

Journal: International Journal of Molecular Sciences

Article Title: Strong Signs for a Weak Wall in Tricuspid Aortic Valve Associated Aneurysms and a Role for Osteopontin in Bicuspid Aortic Valve Associated Aneurysms

doi: 10.3390/ijms20194782

Figure Lengend Snippet: TAV-associated increase in smooth muscle cell density and decrease in α-SM actin expression per smooth muscle cell in situ. ( A ) shows the results of the quantification of smooth muscle cell density in aneurysmal and control media tissue specimens. In ( B ) representative images of aortic tissues following immunofluorescence based detection: wheat germ agglutinin (WGA, cell surface, red), α-SM actin (green), and cell nuclei (TO-PRO-3, blue) are shown. Magnification of images is 63x. In ( C ) the quantification of the amount of Ki67 positive smooth muscle cells is depicted with corresponding representative images shown in ( D ). Magnification of images taken is 63×. ( E ) shows the quantification of α-SM actin expression per smooth muscle cell within the aortic media. (Control n = 30; TAV n = 28; BAV n = 19) In ( F ), the quantification of α-SM actin expression per medial area is shown. (Control n = 30; TAV n = 28; BAV n = 19) In correspondence, ( G ) gives the quantification of the expression of α-SM actin within the isolated smooth muscle cell-lines in vitro . ( H ) shows the results of the quantification of smooth muscle cell-contraction (% of the length of relaxed cells in relation to cell area) ability in response to carbamylcholine chloride in vitro. (Control n = 9; TAV n = 10; BAV n = 10) ( I ) shows representative images of isolated smooth muscle cells stained for α-SM actin (magnification 60×). (** < 0.01).

Article Snippet: Incubation using primary mouse monoclonal anti-α-SM actin antibody (A 2547; Merck Millipore, Darmstadt, Germany) with a dilution of 1:400 was performed overnight at 4 °C.

Techniques: Expressing, In Situ, Control, Immunofluorescence, Isolation, In Vitro, Staining