School of Ophthalmology and Optometry, The Eye Hospital of Wenzhou Medical University, No. 270, Xueyuan Road, Wenzhou, Zhejiang, 325027, China.
Department of Chemical and Materials Engineering, National Central University, No. 300, Jhongda RD., Jhongli, Taoyuan, 32001, Taiwan.
J Mater Chem B. 2021 Sep 29;9(37):7662-7673. doi: 10.1039/d1tb01555a.
We developed poly(vinyl alcohol--itaconic acid) (PV) hydrogels grafted with laminin-derived peptides that had different joint segments and several specific designs, including dual chain motifs. PV hydrogels grafted with a peptide derived from laminin-β4 (PMQKMRGDVFSP) containing a joint segment, dual chain motif and cationic amino acid insertion could attach human pluripotent stem (hPS) cells and promoted high expansion folds in long-term culture (over 10 passages) with low differentiation rates, whereas hPS cells attached poorly on PV hydrogels grafted with laminin-α5 peptides that had joint segments with and without a cationic amino acid or on PV hydrogels grafted with laminin-β4 peptides containing the joint segment only. The inclusion of a cationic amino acid in the laminin-β4 peptide was critical for hPS cell attachment on PV hydrogels, which contributed to the zeta potential shifting to higher values (3-4 mV enhancement). The novel peptide segment-grafted PV hydrogels developed in this study supported hPS cell proliferation, which induced better hPS cell expansion than recombinant vitronectin-coated dishes (gold standard of hPS cell culture dishes) in xeno-free culture conditions. After long-term culture on peptide-grafted hydrogels, hPS cells could be induced to differentiate into specific lineages of cells, such as cardiomyocytes, with high efficiency.
我们开发了接枝有层粘连蛋白衍生肽的聚(乙烯醇-衣康酸)(PV)水凝胶,这些肽具有不同的连接段和几种特定的设计,包括双链基序。接枝有层粘连蛋白-β4 衍生肽(PMQKMRGDVFSP)的 PV 水凝胶,其中包含一个连接段、双链基序和阳离子氨基酸插入,可以附着人类多能干细胞(hPS),并在长期培养(超过 10 代)中以低分化率促进高扩展折叠,而 hPS 细胞在接枝有层粘连蛋白-α5 肽的 PV 水凝胶上附着不良,这些肽具有带和不带阳离子氨基酸的连接段,或在接枝有仅包含连接段的层粘连蛋白-β4 肽的 PV 水凝胶上附着不良。在层粘连蛋白-β4 肽中包含阳离子氨基酸对于 hPS 细胞在 PV 水凝胶上的附着至关重要,这有助于 zeta 电位向更高值(提高 3-4 mV)转移。本研究中开发的新型肽段接枝 PV 水凝胶支持 hPS 细胞增殖,在无动物源培养条件下,比重组 vitronectin 包被的培养皿(hPS 细胞培养皿的金标准)更能诱导 hPS 细胞的扩增。在肽接枝水凝胶上进行长期培养后,hPS 细胞可以被诱导分化为特定的细胞谱系,如心肌细胞,效率很高。