A self-assembling peptide matrix used to control stiffness and binding site density supports the formation of microvascular networks in three dimensions.
A self-assembling peptide matrix used to control stiffness and binding site density supports the formation of microvascular networks in three dimensions.
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DOI：
10.1016/j.actbio.2013.04.002
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发表时间：
2013-08
影响因子：
9.7
通讯作者：
Gooch, K. J.
中科院分区：
文献类型：
--
作者：
Stevenson, M. D.;Piristine, H.;Hogrebe, N. J.;Nocera, T. M.;Boehm, M. W.;Reen, R. K.;Koelling, K. W.;Agarwal, G.;Sarang-Sieminski, A. L.;Gooch, K. J.
关键词：
一个三维(3-D)细胞培养系统，允许控制底物硬度和整合素结合密度被创建和表征。该系统由两个自组装肽(SAP)序列组成，它们以不同的比例混合，以获得所需的凝胶硬度和粘附性。所使用的特异肽是KFE((乙酰基)-FKFEFKFE-CONH2)和KFE-RGD((乙酰基)-GRGDSP-GG-FKFFKFE-CONH2)，KFE-RGD((乙酰基)-GRGDSP-GG-FKFEFKFE-CONH2)是一个类似的序列，包含RGD整合素结合位点。根据其组成和浓度的不同，这些凝胶的存储模数在~60到6000 pA之间。原子力显微镜显示了由纯KFE和纯KFE-RGD以及这两种多肽的混合物组成的凝胶的类ECM纤维微结构。利用该系统研究基质刚性和粘附性对内皮细胞微血管网络形成和人骨髓间充质干细胞形态的影响。当内皮细胞被包裹在没有结合位点的3D凝胶基质中时，无论其硬度如何，几乎没有细胞伸长和网络形成。相反，含有RGD结合位点的基质促进了健壮的MVN的形成，并且这种MVN的形成程度与基质硬度成反比。与没有结合位点的刚性相同的基质相比，含有RGD功能化多肽的基质的平均网络结构长度增加了约2.5倍，作为MVN形成的定量指标。当RGD存在时，含有hMSC的基质在较高的刚度下促进了细胞投射的数量和长度的增加，但当RGD不存在时，在每个刚性处诱导了圆形的形态。综上所述，这些结果证明了在3-D细胞填充凝胶中同时控制底物硬度和结合部位密度的能力，并揭示了硬度和粘附性对细胞类型特异性的细胞行为的重要作用。
A three-dimensional (3-D) cell culture system that allows control of both substrate stiffness and integrin binding density was created and characterized. This system consisted of two self-assembling peptide (SAP) sequences that were mixed in different ratios to achieve the desired gel stiffness and adhesiveness. The specific peptides used were KFE ((acetyl)-FKFEFKFE-CONH2), which has previously been reported not to support cell adhesion or MVN formation, and KFE-RGD ((acetyl)-GRGDSP-GG-FKFEFKFE-CONH2), which is a similar sequence that incorporates the RGD integrin binding site. Storage modulus for these gels ranged from ~60 to 6000 Pa, depending on their composition and concentration. Atomic force microscopy revealed ECM-like fiber microarchitecture of gels consisting of both pure KFE and pure KFE-RGD as well as mixtures of the two peptides. This system was used to study the contributions of both matrix stiffness and adhesiveness on microvascular network (MVN) formation of endothelial cells and the morphology of human mesenchymal stem cells (hMSC). When endothelial cells were encapsulated within 3-D gel matrices without binding sites, little cell elongation and no network formation occurred, regardless of the stiffness. In contrast, matrices containing the RGD binding site facilitated robust MVN formation, and the extent of this MVN formation was inversely proportional to matrix stiffness. Compared with a matrix of the same stiffness with no binding sites, a matrix containing RGD-functionalized peptides resulted in a ~2.5-fold increase in the average length of network structure, which was used as a quantitative measure of MVN formation. Matrices with hMSC facilitated an increased number and length of cellular projections at higher stiffness when RGD was present, but induced a round morphology at every stiffness when RGD was absent. Taken together, these results demonstrate the ability to control both substrate stiffness and binding site density within 3-D cell-populated gels and reveal an important role for both stiffness and adhesion on cellular behavior that is cell-type specific.
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影响因子：
3.7
作者：
Kotlarchyk MA;Shreim SG;Alvarez-Elizondo MB;Estrada LC;Singh R;Valdevit L;Kniazeva E;Gratton E;Putnam AJ;Botvinick EL
通讯作者：
Botvinick EL
多组分自组装生物材料的定向混合。
影响因子：
6.2
作者：
Gasiorowski JZ;Collier JH
通讯作者：
Collier JH
影响因子：
56.9
作者：
Cukierman, E;Pankov, R;Yamada, KM
通讯作者：
Yamada, KM
影响因子：
3.4
作者：
Engler, A;Bacakova, L;Discher, D
通讯作者：
Discher, D
DOI：
10.1016/j.bbrc.2010.09.052
发表时间：
2010-10-15
影响因子：
3.1
作者：
Leong, Wen Shing;Tay, Chor Yong;Tan, Lay Poh
通讯作者：
Tan, Lay Poh