Combinatorial extracellular matrix microenvironments promote survival and phenotype of human induced pluripotent stem cell-derived endothelial cells in hypoxia.
Combinatorial extracellular matrix microenvironments promote survival and phenotype of human induced pluripotent stem cell-derived endothelial cells in hypoxia.
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DOI：
10.1016/j.actbio.2016.08.003
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发表时间：
2016-10-15
影响因子：
9.7
通讯作者：
Huang, Ngan F.
中科院分区：
文献类型：
--
作者：
Hou, Luqia;CoIler, John;Natu, Vanita;Hastie, Trevor J.;Huang, Ngan F.
使用人诱导多能干细胞衍生的内皮细胞（iPSC-ECs）的细胞疗法的最新发展为治疗缺血性心血管组织带来了巨大希望。然而，移植后存活能力差在很大程度上限制了干细胞治疗的潜力。尽管细胞外基质（ECM）已被越来越多地认为是重要的细胞存活因子，但常规方法主要依赖于单个ECM与细胞的体内共递送，即使内皮基底膜由不同ECM的环境组成。为了解决这一限制，我们开发了一种组合ECM微阵列平台，以同时询问ECM的数百种微尺度多组分化学组合物对iPSC-EC响应的影响。在将iPSC-EC接种到ECM微阵列上之后，我们进行了组合ECM对iPSC-EC存活、内皮表型和一氧化氮产生的影响的高通量分析，所述iPSC-EC存活、内皮表型和一氧化氮产生在缺氧（1%O2）和减少的营养物（1%胎牛血清）的条件下，如在缺血性损伤部位中存在的那样。使用自动化图像采集和分析，我们确定了组合ECM，如胶原IV+明胶+硫酸乙酰肝素+层粘连蛋白和胶原IV+纤连蛋白+明胶+硫酸乙酰肝素+层粘连蛋白，显着提高细胞存活率，一氧化氮的产生，和CD 31表型表达，相比于单组分ECM。这些结果在常规细胞培养平台和三维支架内得到进一步验证。此外，这种方法揭示了复杂的ECM相互作用和非直观的细胞行为，否则使用常规细胞培养平台无法容易地确定。总之，这些数据表明，在最佳组合ECM内的iPSC-EC递送可以改善它们在营养物减少的缺氧条件下的存活和功能。
Recent developments in cell therapy using human induced pluripotent stem cell-derived endothelial cells (iPSC-ECs) hold great promise for treating ischemic cardiovascular tissues. However, poor post-transplantation viability largely limits the potential of stem cell therapy. Although the extracellular matrix (ECM) has become increasingly recognized as an important cell survival factor, conventional approaches primarily rely on single ECMs for in vivo co-delivery with cells, even though the endothelial basement membrane is comprised of a milieu of different ECMs. To address this limitation, we developed a combinatorial ECM microarray platform to simultaneously interrogate hundreds of micro-scale multi-component chemical compositions of ECMs on iPSC-EC response. After seeding iPSC-ECs onto ECM microarrays, we performed high-throughput analysis of the effects of combinatorial ECMs on iPSC-EC survival, endothelial phenotype, and nitric oxide production under conditions of hypoxia (1% O2) and reduced nutrients (1% fetal bovine serum), as is present in ischemic injury sites. Using automated image acquisition and analysis, we identified combinatorial ECMs such as collagen IV+gelatin+heparan sulfate+laminin and collagen IV+fibronectin+gelatin+heparan sulfate+laminin that significantly improved cell survival, nitric oxide production, and CD31 phenotypic expression, in comparison to single-component ECMs. These results were further validated in conventional cell culture platforms and within three-dimensional scaffolds. Furthermore, this approach revealed complex ECM interactions and non-intuitive cell behavior that otherwise could not be easily determined using conventional cell culture platforms. Together these data suggested that iPSC-EC delivery within optimal combinatorial ECMs may improve their survival and function under the condition of hypoxia with reduced nutrients.
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影响因子：
56.9
作者：
Asahara, T;Murohara, T;Isner, JM
通讯作者：
Isner, JM
DOI：
10.1073/pnas.76.10.5217
发表时间：
1979-01-01
影响因子：
11.1
作者：
FOLKMAN, J;HAUDENSCHILD, CC;ZETTER, BR
通讯作者：
ZETTER, BR
影响因子：
14.8
作者：
Brafman, David A.;Chien, Shu;Willert, Karl
通讯作者：
Willert, Karl
DOI：
10.1083/jcb.200405004
发表时间：
2004-09-13
期刊：
The Journal of cell biology
影响因子：
--
作者：
Engler AJ;Griffin MA;Sen S;Bönnemann CG;Sweeney HL;Discher DE
通讯作者：
Discher DE
影响因子：
14
作者：
Brafman, David A.;Chang, Chien W.;Fernandez, Antonio;Willert, Karl;Varghese, Shyni;Chien, Shu
通讯作者：
Chien, Shu