Cardiac differentiation of induced pluripotent stem cells on elastin-like protein-based hydrogels presenting a single-cell adhesion sequence
Cardiac differentiation of induced pluripotent stem cells on elastin-like protein-based hydrogels presenting a single-cell adhesion sequence
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诱导多能干细胞在弹性蛋白样蛋白水凝胶上的心脏分化呈现单细胞粘附序列
DOI：
10.1038/s41428-018-0110-2
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发表时间：
2019
期刊：
影响因子：
2.8
通讯作者：
Kambe Y，Tokushige T，Mahara A，Iwasaki Y，Yamaoka T
中科院分区：
文献类型：
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作者：
Kambe Y，Tokushige T，Mahara A，Iwasaki Y，Yamaoka T
在不同的细胞外基质(ECM)来源的底物上，如I型胶原(Col-I)，研究了诱导多能干细胞(IPSCs)向心脏分化的底物依赖性。然而，ECM来源的底物具有多种细胞黏附氨基酸序列，刺激细胞内各种信号通路，使人们很难阐明底物依赖的干细胞分化的机制。呈现这些序列之一的底物是阐明这一机制的有力工具。我们设计了由带有或不带有RGD细胞黏附基序的重复VPGIG序列组成的弹性蛋白样蛋白(ELP-RGD/ELP-Ctrl)，并使用化学交联剂产生水凝胶。通过调节ELP和交联剂的浓度，我们得到了杨氏模数为0.3kpa的ELP-Ctrl和ELP-RGD型水凝胶。用ELP-Ctrl和ELP-RGD凝胶作为培养的小鼠IPSCs向心脏分化的底物。在ELP-RGD凝胶上的细胞表现出比在Col-I凝胶上高四倍的收缩蛋白T型肌钙蛋白的基因表达，Col-I凝胶是IPSC心脏分化的有效底物。ELP-RGD凝胶可能会刺激细胞中整合素衍生的信号通路，以促进心脏分化。这项研究显示了ELP水凝胶通过控制细胞黏附序列呈现来研究底物依赖的IPSC心脏分化的潜力。
Substrate-dependent cardiac differentiation of induced pluripotent stem cells (iPSCs) has been studied on various extracellular matrix (ECM)-derived substrates, such as collagen type I (Col-I). However, ECM-derived substrates have multiple cell-adhesive amino acid sequences and stimulate various signaling pathways in cells, making it difficult to clarify the mechanism of substrate-dependent stem cell differentiation. A substrate presenting one of these sequences is a powerful tool for elucidating the mechanism. We designed elastin-like proteins (ELPs) composed of repetitive VPGIG sequences with or without the RGD cell adhesion motif (ELP-RGD/ELP-Ctrl) and used a chemical crosslinker to generate hydrogels. By adjusting the ELP and crosslinker concentrations, we obtained ELP-Ctrl and ELP-RGD hydrogels with a Young’s modulus of 0.3 kPa. The ELP-Ctrl and ELP-RGD gels were used as a substrate for the cardiac differentiation of cultured murine iPSCs. Cells on the ELP-RGD gel showed four times higher gene expression of the contractile protein troponin T type 2 than those on a Col-I gel, which is an effective substrate for iPSC cardiac differentiation. The ELP-RGD gel might stimulate integrin-derived signaling pathways in the cells to promote cardiac differentiation. This study showed the potential of ELP hydrogels for studying substrate-dependent iPSC cardiac differentiation by enabling the control of cell-adhesive sequence presentation.