Hyperbranched poly(ϵ-lysine) substrate presenting the laminin sequence YIGSR induces the formation of spheroids in adult bone marrow stem cells.
Hyperbranched poly(ϵ-lysine) substrate presenting the laminin sequence YIGSR induces the formation of spheroids in adult bone marrow stem cells.
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DOI：
10.1371/journal.pone.0187182
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Santin M
中科院分区：
文献类型：
--
作者：
Perugini V;Meikle ST;Guildford AL;Santin M
与人间充质干细胞单层培养后形成的成纤维细胞样细胞不同，骨髓和其他类型组织的天然干细胞龛有利于形成三维（3D）细胞簇。这些簇的结构和生物活性由细胞与基底膜和相邻细胞建立的接触来调节，并导致它们的不对称分裂和随后的干细胞群和定向后代的维持。目前的工作证明了合成底物在体外模拟干细胞龛的潜力。线性聚-L-赖氨酸的侧氨基用超支化聚-（β-赖氨酸）肽修饰，称为树枝状分子，与层粘连蛋白模拟序列YIGSR连接。这些树突在最高分子分支处呈现YIGSR序列，确保生物配体的受控间距。当用于包被无血清体外细胞培养系统中的组织培养板的表面时，基底能够模拟干细胞龛的基底膜的最相关特征，即IV型胶原的网状结构和与整联蛋白生物识别相关的层粘连蛋白生物配体的可用性。测试了基质仿生性质支持形成与天然干细胞龛中观察到的类似的人骨髓间充质干细胞（hMSC）3D球状体的能力以及维持干细胞多能性标记物的能力。这些特征与以下物质的底物特异性表达和定位相关：（i）细胞粘附受体（即β-整联蛋白和N-钙粘蛋白），（ii）多能性标志物和细胞骨架蛋白的转录因子，以及（iii）整个细胞培养物第2 - 4代细胞迁移的调节因子。结果清楚地表明，从基质粘附扩散细胞的不对称分裂开始的3D球状体的形成，相关整合素的聚集和控制细胞骨架形成的特定细胞内途径的表达，表明它们可能用作移植程序前处理干细胞的基质。
Unlike the fibroblast-like cells formed upon monolayer culture of human mesenchymal stem cells, the natural stem cell niche of the bone marrow and other types of tissues favours the formation of 3-dimensional (3D) cell clusters. The structuring and biological activity of these clusters are regulated by the contacts established by cells with both the basement membrane and neighbour cells and results in their asymmetric division and the consequent maintenance of both a stem population and a committed progeny. The present work demonstrates the potential of a synthetic substrate to mimic the stem cell niche in vitro. The side amino groups of a linear Poly-L-lysine were modified with hyperbranched poly-(ϵ-lysine) peptides, named as dendrons, tethered with the laminin-mimicking sequence, YIGSR. These dendrons presented the YIGSR sequence at the uppermost molecular branching ensuring a controlled spacing of the bioligand. When used to coat the surface of tissue culture plates in a serum-free in vitro cell culture system, the substrate was able to mimic the most relevant features of the basement membrane of the stem cell niche, i.e. the mesh structure of Collagen Type IV and the availability of laminin bioligands relevant to integrin biorecognition. The substrate biomimetic properties were tested for their ability to support the formation of human bone marrow mesenchymal stem cells (hMSCs) 3D spheroids similar to those observed in the natural stem cell niches and their ability to maintain stem cell pluripotency markers. These features were related to the substrate-specific expression and localisation of (i) cell adhesion receptors (i.e. β-integrin and N-cadherin), (ii) transcription factors of pluripotency markers and cytoskeleton protein and (iii) regulators of cell migration throughout cell culture passages 2 to 4. The results clearly demonstrate the formation of 3D spheroids starting from the asymmetric division of substrate-adhering spread cells, the clustering of relevant integrins and the expression of specific intracellular pathways controlling cytoskeleton formation suggesting their potential use as a substrate for the handling of stem cells prior to transplantation procedures.
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影响因子：
7.4
作者：
Liras A
通讯作者：
Liras A
DOI：
10.1089/ten.teb.2009.0270
发表时间：
2009-09
期刊：
Tissue engineering. Part B, Reviews
影响因子：
--
作者：
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DOI：
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发表时间：
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影响因子：
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影响因子：
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