A feeder-free, defined three-dimensional polyethylene glycol-based extracellular matrix niche for culture of human embryonic stem cells
A feeder-free, defined three-dimensional polyethylene glycol-based extracellular matrix niche for culture of human embryonic stem cells
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DOI：
10.1016/j.biomaterials.2013.01.073
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发表时间：
2013-05-01
期刊：
影响因子：
14
通讯作者：
Lim, Jeong Mook
中科院分区：
文献类型：
--
作者：
Jang, Mi;Lee, Seung Tae;Lim, Jeong Mook
我们报告了一个无血清和饲养层的优化，三维（3D）的小生境与合成的聚乙二醇（PEG）为基础的细胞外基质的自我更新的人胚胎干细胞（hESC）。三个hESC系（H9，H1和Novo）在不同的机械性能的水凝胶中培养，并将细胞形态和活性与无饲养层或含饲养层的二维（2D）小生境中的培养进行比较。在H9细胞系中检测到PEG浓度（5%、7.5%、10%、12.5%或15%）和乙烯砜官能化的PEG多臂数（3%、4%或8%）对hESC形态的显著影响。任何PEG浓度的8多臂结构的细胞生长最大，这产生了干细胞相关基因的最高表达。培养的H9细胞中的碱性磷酸酶活性在优化的无饲养层的3D和含饲养层的2D系统之间是相似的。然而，在3D培养的hESC团块中检测到KLF 4、CDH 1、TERT、SOX 2和UTF 1基因的表达增加以及多能特异性SSEA-4、Oct 3/4、Nanog、Tra-1-60和Tra-1-81的表达。H1和Novo细胞系也在优化的3D系统中扩增，其保持了干性特性。虽然三个品系的增殖活性不同，但经过三维培养后，差异有所减小。这些结果表明，使用PEG基水凝胶产生的化学定义的非细胞小生境具有支持hESC自我更新的潜力。3D特性的调制可以创建用于细胞转化和分化的各种模型。(C)2013爱思唯尔有限公司版权所有。
We report optimization of a serum- and feeder-free, three-dimensional (3D) niche created with a synthetic polyethylene glycol (PEG)-based extracellular matrix for self-renewal of human embryonic stem cells (hESCs). Three hESC lines (H9, H1 and Novo) were cultured in hydrogels of different mechanical properties, and cellular morphology and activity were compared to culture in feeder-free or feeder-containing two-dimensional (2D) niches. Significant effects of PEG concentration (5, 7.5, 10, 12.5 or 15%) and vinyl sulfone-functionalized PEG multiarm number (3, 4 or 8) on hESC morphology were detected in the H9 line. Cell growth was maximal with an 8 multiarm architecture of any PEG concentration, which yielded the highest expression of sternness-related genes. Alkaline phosphatase activity in cultured H9 cells was similar between the optimized feeder-free 3D and the feeder-containing 2D systems. However, increased expression of the KLF4, CDH1, TERT, SOX2, and UTF1 genes and expression of pluripotency-specific SSEA-4, Oct3/4, Nanog, Tra-1-60 and Tra-1-81 were detected in the 3D-cultured hESC clumps. H1 and Novo cell lines also expanded in the optimized 3D system, which maintain sternness properties. Although different proliferation activities were detected among three lines, the difference was decreased after the 3D culture. These results demonstrate that chemically defined, acellular niches created using PEG-based hydrogels have the potential to support hESC self-renewal. Modulation of 3D properties can create various models for cell transformation and differentiation. (C) 2013 Elsevier Ltd. All rights reserved.