E dimensional (3D) matrixes (Fig. 1), [9, 37?9] which is supported by the fact > 자유게시판

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E dimensional (3D) matrixes (Fig. 1), [9, 37?9] which is supported by the fact that HUVECs cultured without another cell type did not form any organized structures. After 14 days of coculturing HUVECs with BMSCs, the expression of alpha smooth muscle actin (-SMA) in BMSCs increased compared to expression before Lenvatinib co-culture and surrounded EC cord-like structures. A multiplex chemiluminescent ELISA was performed by Verseijden et al. after 14 days of BMSC monoculture to analyze which angiogenic factors were increased in BMSC-conditioned medium compared to unconditioned medium or HUVEC-conditionedmedium [9]. Results showed significantly higher amounts of hepatocyte growth factor (HGF), tissue inhibitor of metalloproteinase 1, and tissue inhibitor of metalloproteinase 2 (TIMP1 and TIMP2) in co-culture compared to monoculture. These factors are known also to regulate vessel formation. Additionally, HGF stimulates EC proliferation and induces vessel formation [40]. On the other hand, experiments using different concentrations of HGF failed to induce HUVEC outgrowth in monocultures. Regarding the high amount of TIMPs, further investigations have been performed because TIMPs had also been reported to inhibit the activity of matrix metalloproteinases (MMPs) [41]. Because of the high concentration of serum in the growth media, which is known PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/12711626 to have a high MMP concentration itself, it was not possible to discriminate the amount of MMPs secreted specifically by BMSCs. Nevertheless, much of the fibrin was removed within the fibrin matrix, suggesting that MMPs or other factors were secreted in sufficient amount to degrade the fibrin. Despite the findings of angiogenesis related factors in BMSC-conditioned medium, HUVECs treated with this medium showed no positive effect regarding outgrowth or organization. The fact that BMSCs co-cultured with HUVECs induced outgrowth showed that HUVEC outgrowth and organization into vessel-like structures is not merely achieved by secreted factors. This suggests that direct cell-cell contact and reciprocal signaling may play an important role for ECs to form pre-vascular-like structures [9]. To investigate the angiogenesis promoting and vessel stabilization functions of BMSCs on ECs, BMSCs have been co-cultured with EPCs or HUVECs in a 3D polyurethane scaffold [37]. Luminal tubular structures were detected after 7 days of culture, which were not only positive for PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/8627573 endothelial cell markers CD31 and vWF, but also for CD146, expressed by pericytes but not ECs. Sequential sectioning of scaffolds revealed areas positive for CD146, neuron-glial antigen 2 (NG2), and -SMA, which were tightly associated with tubular structures. TheFig. 1 Example of how BMSCs support EC network formation in a fibrin matrix. a Fluorescent image of a capillary network on day 7. Thresholds were set before applying the Angiogenesis Tube Formation Application Module in Metamorph imaging software. b To visualize networks, the filament tracing function was used using Imaris. ECs are depicted in blue and BMSCs in red [39]Pill et al. Cell Regeneration (2015) 4:Page 4 ofcombination of CD146, NG2, and -SMA confirmed the pericyte phenotype. Furthermore, neither EPCs nor BMSCs cultured alone could form pre-vascular structures which showed the major role of BMSCs in promoting EPCs to differentiate into a mature network. These experiments further indicated that both cell types influence each other in terms of differentiation. BMSCs did prevent a decrease in nu.