The use of vascular endothelial growth factor functionalized agarose to guide pluripotent stem cell aggregates toward blood progenitor cells
The use of vascular endothelial growth factor functionalized agarose to guide pluripotent stem cell aggregates toward blood progenitor cells
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DOI：
10.1016/j.biomaterials.2010.07.040
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发表时间：
2010-11-01
期刊：
影响因子：
14
通讯作者：
Shoichet, Molly S.
中科院分区：
文献类型：
--
作者：
Rahman, Nafees;Purpura, Kelly A.;Shoichet, Molly S.
多能干细胞的发育潜力受其局部细胞微环境的影响。为了更好地了解血管内皮生长因子(VEGFA)在胚胎细胞微环境中的作用，我们合成了一个人工干细胞壁龛，将VEGFA固定在琼脂糖凝胶中。琼脂糖首先用香豆素保护的硫醇修饰。在紫外线激发下，香豆素基团被裂解，留下活性硫醇与马来酰亚胺激活的VEGFA偶联。将小鼠胚胎干细胞(ESC)聚集体包裹在VEGFA固定化琼脂糖凝胶中，在无血清条件下以自由漂浮的聚集体形式培养7d。对包裹的聚集体进行了造血祖细胞能力的评估。在骨形态发生蛋白-4(BMP-4)存在的情况下，固定化VEGFA处理的细胞在第4天上调中胚层标志物、短暂性生长因子和血管内皮生长因子受体2(T+VEGFR2+)，第7天表达CD34和CD41(CD34+CD41+)。结果发现，固定化VEGFA在每分子基础上诱导造血祖细胞(包括克隆形成细胞)的效率高于可溶性VEGFA。这项工作展示了使用功能化水凝胶引导包裹的ESCs走向血祖细胞，并介绍了一种能够概括胚胎微环境方面的工具。(C)2010爱思唯尔有限公司。保留所有权利。
The developmental potential of pluripotent stem cells is influenced by their local cellular microenvironment. To better understand the role of vascular endothelial growth factor (VEGFA) in the embryonic cellular microenvironment, we synthesized an artificial stem cell niche wherein VEGFA was immobilized in an agarose hydrogel. Agarose was first modified with coumarin-protected thiols. Upon exposure to ultra-violet excitation, the coumarin groups were cleaved leaving reactive thiols to couple with maleimide-activated VEGFA. Mouse embryonic stem cells (ESC) aggregates were encapsulated in VEGFA immobilized agarose and cultured for 7 days as free-floating aggregates under serum-free conditions. Encapsulated aggregates were assessed for their capacity to give rise to blood progenitor cells. In the presence of bone morphogenetic protein-4 (BMP-4), cells exposed to immobilized VEGFA upregulated mesodermal markers, brachyury and VEGF receptor 2 (T+VEGFR2+) by day 4, and expressed CD34 and CD41 (CD34+CD41+) on day 7. It was found that immobilized VEGFA treatment was more efficient at inducing blood progenitors (including colony forming cells) on a per molecule basis than soluble VEGFA. This work demonstrates the use of functionalized hydrogels to guide encapsulated ESCs toward blood progenitor cells and introduces a tool capable of recapitulating aspects of the embryonic microenvironment. (C) 2010 Elsevier Ltd. All rights reserved.