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Interactions of human endothelial and multipotent mesenchymal stem cells in cocultures.

Ern C, Krump-Konvalinkova V, Docheva D, Schindler S, Rossmann O, Böcker W, Mutschler W, Schieker M - Open Biomed Eng J (2010)

Bottom Line: Current strategies for tissue engineering of bone rely on the implantation of scaffolds, colonized with human mesenchymal stem cells (hMSC), into a recipient.A major limitation is the lack of blood vessels.One approach to enhance the scaffold vascularisation is to supply the scaffolds with endothelial cells (EC).The main goal of this study was to establish a coculture system of hMSC and EC for the purposes of bone tissue engineering.

View Article: PubMed Central - PubMed

Affiliation: Experimental Surgery and Regenerative Medicine, Department of Surgery, University of Munich (LMU), Munich, Germany.

ABSTRACT
Current strategies for tissue engineering of bone rely on the implantation of scaffolds, colonized with human mesenchymal stem cells (hMSC), into a recipient. A major limitation is the lack of blood vessels. One approach to enhance the scaffold vascularisation is to supply the scaffolds with endothelial cells (EC).The main goal of this study was to establish a coculture system of hMSC and EC for the purposes of bone tissue engineering. Therefore, the cell behaviour, proliferation and differentiation capacity in various cell culture media as well as cell interactions in the cocultures were evaluated.The differentiation capacity of hMSC along osteogenic, chondrogenic, and adipogenic lineage was impaired in EC medium while in a mixed EC and hMSC media, hMSC maintained osteogenic differentiation. In order to identify and trace EC in the cocultures, EC were transduced with eGFP. Using time-lapse imaging, we observed that hMSC and EC actively migrated towards cells of their own type and formed separate clusters in long term cocultures. The scarcity of hMSC and EC contacts in the cocultures suggest the influence of growth factor-mediated cell interactions and points to the necessity of further optimization of the coculture conditions.

No MeSH data available.


Related in: MedlinePlus

Growth media affect the differentiation potential of hMSC in vitro. The hMSC were grown in M1 (A, D, G), M2 (B, E, H), and M3 (C, F, J) for 21 days, and subsequently subjected to differentiation. The osteogenic differentiation was detected by means of von Kossa staining (A, B, C), adipogenic differentiation by means of oil red O staining (D, E, F), and chondrogenic differentiation by means of collagen II staining (G, H, J). Bar represent 200 µm (B), 100 µm (A, C, G insert), 50 µm (D, E, F), and 300 µm (G, H, J).
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Figure 4: Growth media affect the differentiation potential of hMSC in vitro. The hMSC were grown in M1 (A, D, G), M2 (B, E, H), and M3 (C, F, J) for 21 days, and subsequently subjected to differentiation. The osteogenic differentiation was detected by means of von Kossa staining (A, B, C), adipogenic differentiation by means of oil red O staining (D, E, F), and chondrogenic differentiation by means of collagen II staining (G, H, J). Bar represent 200 µm (B), 100 µm (A, C, G insert), 50 µm (D, E, F), and 300 µm (G, H, J).

Mentions: In order to evaluate the effects of growth media on the differentiation potential of hMSC and to prove their stem cell character, we propagated the hMSC of the three donors in the three growth media for 21 days and subsequently subjected the cells to treatments known to induce osteogenic, chondrogenic, and adipogenic differentiation. Osteogenic differentiation could be induced in hMSC propagated in all three media tested, as shown by means of von Kossa staining (Fig. 4A, B, C). In addition, the expression of the osteogenic differentiation marker osterix in the cells that have been propagated in M1, M2 or M3 was demonstrated by RT-PCR (Fig. 5).


Interactions of human endothelial and multipotent mesenchymal stem cells in cocultures.

Ern C, Krump-Konvalinkova V, Docheva D, Schindler S, Rossmann O, Böcker W, Mutschler W, Schieker M - Open Biomed Eng J (2010)

Growth media affect the differentiation potential of hMSC in vitro. The hMSC were grown in M1 (A, D, G), M2 (B, E, H), and M3 (C, F, J) for 21 days, and subsequently subjected to differentiation. The osteogenic differentiation was detected by means of von Kossa staining (A, B, C), adipogenic differentiation by means of oil red O staining (D, E, F), and chondrogenic differentiation by means of collagen II staining (G, H, J). Bar represent 200 µm (B), 100 µm (A, C, G insert), 50 µm (D, E, F), and 300 µm (G, H, J).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3102321&req=5

Figure 4: Growth media affect the differentiation potential of hMSC in vitro. The hMSC were grown in M1 (A, D, G), M2 (B, E, H), and M3 (C, F, J) for 21 days, and subsequently subjected to differentiation. The osteogenic differentiation was detected by means of von Kossa staining (A, B, C), adipogenic differentiation by means of oil red O staining (D, E, F), and chondrogenic differentiation by means of collagen II staining (G, H, J). Bar represent 200 µm (B), 100 µm (A, C, G insert), 50 µm (D, E, F), and 300 µm (G, H, J).
Mentions: In order to evaluate the effects of growth media on the differentiation potential of hMSC and to prove their stem cell character, we propagated the hMSC of the three donors in the three growth media for 21 days and subsequently subjected the cells to treatments known to induce osteogenic, chondrogenic, and adipogenic differentiation. Osteogenic differentiation could be induced in hMSC propagated in all three media tested, as shown by means of von Kossa staining (Fig. 4A, B, C). In addition, the expression of the osteogenic differentiation marker osterix in the cells that have been propagated in M1, M2 or M3 was demonstrated by RT-PCR (Fig. 5).

Bottom Line: Current strategies for tissue engineering of bone rely on the implantation of scaffolds, colonized with human mesenchymal stem cells (hMSC), into a recipient.A major limitation is the lack of blood vessels.One approach to enhance the scaffold vascularisation is to supply the scaffolds with endothelial cells (EC).The main goal of this study was to establish a coculture system of hMSC and EC for the purposes of bone tissue engineering.

View Article: PubMed Central - PubMed

Affiliation: Experimental Surgery and Regenerative Medicine, Department of Surgery, University of Munich (LMU), Munich, Germany.

ABSTRACT
Current strategies for tissue engineering of bone rely on the implantation of scaffolds, colonized with human mesenchymal stem cells (hMSC), into a recipient. A major limitation is the lack of blood vessels. One approach to enhance the scaffold vascularisation is to supply the scaffolds with endothelial cells (EC).The main goal of this study was to establish a coculture system of hMSC and EC for the purposes of bone tissue engineering. Therefore, the cell behaviour, proliferation and differentiation capacity in various cell culture media as well as cell interactions in the cocultures were evaluated.The differentiation capacity of hMSC along osteogenic, chondrogenic, and adipogenic lineage was impaired in EC medium while in a mixed EC and hMSC media, hMSC maintained osteogenic differentiation. In order to identify and trace EC in the cocultures, EC were transduced with eGFP. Using time-lapse imaging, we observed that hMSC and EC actively migrated towards cells of their own type and formed separate clusters in long term cocultures. The scarcity of hMSC and EC contacts in the cocultures suggest the influence of growth factor-mediated cell interactions and points to the necessity of further optimization of the coculture conditions.

No MeSH data available.


Related in: MedlinePlus