Differentiation of Induced Pluripotent Stem Cells towards Mesenchymal Stromal Cells is Hampered by Culture in 3D Hydrogels
Differentiation of Induced Pluripotent Stem Cells towards Mesenchymal Stromal Cells is Hampered by Culture in 3D Hydrogels
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DOI：
10.1038/s41598-019-51911-5
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发表时间：
2019-10-30
影响因子：
4.6
通讯作者：
Wagner, Wolfgang
中科院分区：
文献类型：
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作者：
Goetzke, Roman;Keijdener, Hans;Wagner, Wolfgang
诱导多能干细胞（iPSC）向特定谱系的定向分化仍然是再生医学的主要挑战，同时越来越多的人认为这一过程可能受到三维环境的影响。在这项研究中，我们研究了iPSCs是否可以向间充质基质细胞（MSC）分化时，嵌入纤维蛋白水凝胶，使一个一步分化过程中的支架。iPSC在组织培养塑料或纤维蛋白水凝胶上的分化导致典型的MSC样表型。相比之下，包埋到纤维蛋白凝胶中的iPSC产生了小得多的细胞，其具有异质性生长模式，不存在纤连蛋白，微弱的CD73和CD105表达，以及向成骨和成脂谱系的分化潜力降低。转录组学分析表明，MSC和细胞外基质的特征基因在平面基板上上调，而神经发育的基因在三维培养中上调。此外，3D培养对DNA甲基化谱有重大影响，特别是在神经元和心血管发育的基因中，而没有证据表明MSC的表观遗传成熟。总之，iPSC可以在组织培养塑料或平坦的纤维蛋白水凝胶上向MSC分化。相反，当iPSC嵌入水凝胶中时，分化过程是异质的，并且不针对MSC。
Directed differentiation of induced pluripotent stem cells (iPSCs) towards specific lineages remains a major challenge in regenerative medicine, while there is a growing perception that this process can be influenced by the three-dimensional environment. In this study, we investigated whether iPSCs can differentiate towards mesenchymal stromal cells (MSCs) when embedded into fibrin hydrogels to enable a one-step differentiation procedure within a scaffold. Differentiation of iPSCs on tissue culture plastic or on top of fibrin hydrogels resulted in a typical MSC-like phenotype. In contrast, iPSCs embedded into fibrin gel gave rise to much smaller cells with heterogeneous growth patterns, absence of fibronectin, faint expression of CD73 and CD105, and reduced differentiation potential towards osteogenic and adipogenic lineage. Transcriptomic analysis demonstrated that characteristic genes for MSCs and extracellular matrix were upregulated on flat substrates, whereas genes of neural development were upregulated in 3D culture. Furthermore, the 3D culture had major effects on DNA methylation profiles, particularly within genes for neuronal and cardiovascular development, while there was no evidence for epigenetic maturation towards MSCs. Taken together, iPSCs could be differentiated towards MSCs on tissue culture plastic or on a flat fibrin hydrogel. In contrast, the differentiation process was heterogeneous and not directed towards MSCs when iPSCs were embedded into the hydrogel.