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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

The proliferation rate of hMSC is donor dependent. Cumulative population doublings of hMSC from 3 different donors (A, B, C) and AS-M.5 (D) in cultures supplemented with M1 (diamonds, full line), M3 (triangles, dashed), and M2 (squares, dashed line) in the course of 21 days of cultivation. Indicated are means +/- SEM.
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Figure 2: The proliferation rate of hMSC is donor dependent. Cumulative population doublings of hMSC from 3 different donors (A, B, C) and AS-M.5 (D) in cultures supplemented with M1 (diamonds, full line), M3 (triangles, dashed), and M2 (squares, dashed line) in the course of 21 days of cultivation. Indicated are means +/- SEM.

Mentions: Major differences in the proliferation rate of hMSC isolated from three different donors were observed in the three media tested. The cumulative population doubling of hMSC in the course of 21 days of cultivation have shown significant donor-dependent differences (Fig. 2). While hMSC of donor II and III (Fig. 2B, C) proliferated in M3 more rapidly than in M1 during the first two weeks, the growth of hMSC of donor I was in M3 much slower than in the M1. The hMSC of all 3 donors proliferated in M2 either as fast as in the M1 or faster. Similarly, virtually no proliferation of EC was observed in M1 but a slow proliferation of EC was measured in the M2. Even though the cultures of hMSC isolated from 3 different donors proliferated continuously in the M1 in the course of the observation period of 21 days, they differed substantially in the number of population doubling they ultimately reached. While the cultures of donor III reached population doubling of 10, the cultures of the remaining donors did not surpass 7 population doublings. The hMSC cultures isolated from different donors differed also in their capacity to proliferate in M3. While hMSC cultures from donor I and III ceased to proliferate exponentially in M3 after about 2 weeks in culture, the cultures from the donor II continued to proliferate in M3 at a similar rate as in the M1.


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)

The proliferation rate of hMSC is donor dependent. Cumulative population doublings of hMSC from 3 different donors (A, B, C) and AS-M.5 (D) in cultures supplemented with M1 (diamonds, full line), M3 (triangles, dashed), and M2 (squares, dashed line) in the course of 21 days of cultivation. Indicated are means +/- SEM.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: The proliferation rate of hMSC is donor dependent. Cumulative population doublings of hMSC from 3 different donors (A, B, C) and AS-M.5 (D) in cultures supplemented with M1 (diamonds, full line), M3 (triangles, dashed), and M2 (squares, dashed line) in the course of 21 days of cultivation. Indicated are means +/- SEM.
Mentions: Major differences in the proliferation rate of hMSC isolated from three different donors were observed in the three media tested. The cumulative population doubling of hMSC in the course of 21 days of cultivation have shown significant donor-dependent differences (Fig. 2). While hMSC of donor II and III (Fig. 2B, C) proliferated in M3 more rapidly than in M1 during the first two weeks, the growth of hMSC of donor I was in M3 much slower than in the M1. The hMSC of all 3 donors proliferated in M2 either as fast as in the M1 or faster. Similarly, virtually no proliferation of EC was observed in M1 but a slow proliferation of EC was measured in the M2. Even though the cultures of hMSC isolated from 3 different donors proliferated continuously in the M1 in the course of the observation period of 21 days, they differed substantially in the number of population doubling they ultimately reached. While the cultures of donor III reached population doubling of 10, the cultures of the remaining donors did not surpass 7 population doublings. The hMSC cultures isolated from different donors differed also in their capacity to proliferate in M3. While hMSC cultures from donor I and III ceased to proliferate exponentially in M3 after about 2 weeks in culture, the cultures from the donor II continued to proliferate in M3 at a similar rate as in the M1.

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