Matrix composition regulates three-dimensional network formation by endothelial cells and mesenchymal stem cells in collagen/fibrin materials.

Matrix composition regulates three-dimensional network formation by endothelial cells and mesenchymal stem cells in collagen/fibrin materials.
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DOI:
10.1007/s10456-012-9257-1
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发表时间:
2012-06
期刊:
影响因子:
9.8
通讯作者:
Stegemann, Jan P.
Stegemann, Jan P.
中科院分区:
医学1区
文献类型:
--
作者:
Rao, Rameshwar R.;Peterson, Alexis W.;Ceccarelli, Jacob;Putnam, Andrew J.;Stegemann, Jan P.

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内皮细胞(EC)和间充质干细胞(MSC)在三维(3D)蛋白质水凝胶中的共培养物可用于在体外概括血管发生的方面。MSC提供旁分泌信号,刺激EC形成血管样结构,其随着MSC向壁细胞的作用转变而成熟。在这项研究中,血管样网络的形成进行了研究,使用3D胶原/纤维蛋白(COL/FIB)基质接种嵌入EC和MSC和培养7天。EC:MSC比率从5:1、3:2、1:1、2:3和1:5变化。基质组成在COL/FIB组成为100/0(纯COL)、60/40、50/50、40/60和0/100(纯FIB)下变化。相对于其他细胞比率,血管生成在最高EC:MSC比率中显著降低。尽管40/60 COL/FIB和纯纤维蛋白材料表现出相同程度的血管生成,但随着复合材料中纤维蛋白含量的增加,网络形成增加。7天后EC和MSC共定位于血管样结构中,总细胞数增加约70%。力学性能的测量结果表明,矩阵刚度和网络形成之间的负相关性。使用不同总蛋白质含量制备的凝胶和通过使用二醛乙二醛交联基质进一步研究基质刚度的影响。这一系列系统的研究表明,基质组成调节3D蛋白质水凝胶中的血管发生,并进一步表明这种效应可能是由基质机械性能引起的。这些发现与研究新血管形成和发展促进移植组织血管化的策略有关。
Co-cultures of endothelial cells (EC) and mesenchymal stem cells (MSC) in three-dimensional (3D) protein hydrogels can be used to recapitulate aspects of vasculogenesis in vitro. MSC provide paracrine signals that stimulate EC to form vessel-like structures, which mature as the MSC transition to the role of mural cells. In this study, vessel-like network formation was studied using 3D collagen/fibrin (COL/FIB) matrices seeded with embedded EC and MSC and cultured for 7 days. The EC:MSC ratio was varied from 5:1, 3:2, 1:1, 2:3 and 1:5. The matrix composition was varied at COL/FIB compositions of 100/0 (pure COL), 60/40, 50/50, 40/60 and 0/100 (pure FIB). Vasculogenesis was markedly decreased in the highest EC:MSC ratio, relative to the other cell ratios. Network formation increased with increasing fibrin content in composite materials, although the 40/60 COL/FIB and pure fibrin materials exhibited the same degree of vasculogenesis. EC and MSC were co-localized in vessel-like structures after 7 days and total cell number increased by approximately 70%. Mechanical property measurements showed an inverse correlation between matrix stiffness and network formation. The effect of matrix stiffness was further investigated using gels made with varying total protein content and by crosslinking the matrix using the dialdehyde glyoxal. This systematic series of studies demonstrates that matrix composition regulates vasculogenesis in 3D protein hydrogels, and further suggests that this effect may be caused by matrix mechanical properties. These findings have relevance to the study of neovessel formation and the development of strategies to promote vascularization in transplanted tissues.
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