Effect of substrate stiffness on early human embryonic stem cell differentiation.
Effect of substrate stiffness on early human embryonic stem cell differentiation.
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DOI：
10.1186/1754-1611-7-7
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发表时间：
2013-03-21
影响因子：
5.6
通讯作者：
Rao RR
中科院分区：
文献类型：
--
作者：
Eroshenko N;Ramachandran R;Yadavalli VK;Rao RR
人类胚胎干细胞（hESC）的多能性和自我更新特性使其成为发育生物学、药理学和再生医学领域的宝贵工具。因此，人们对设计 hESC 增殖和分化策略产生了极大的兴趣。近年来，涉及模拟天然干细胞微环境（化学和物理）的方法受到了广泛关注。同样重要的是，有证据表明细胞还可以感知其微环境的机械特性。在这项研究中，我们通过在不同硬度的柔性聚二甲基硅氧烷（PDMS）上培养 hESC 来测试其接受机械信号以从基质中分化的假设。 PDMS 基底使用现有的商业配方制备，并表征其硬度、表面特性以及细胞附着和增殖的效率。发现在不同的基底硬度下，细胞数量、细胞附着和细胞表面积是相似的。在不同硬度的所有 PDMS 基质上，多能性标记的表达随着培养时间的增加而减少。分化标记基因表达的分析表明，随着培养时间的延长，分化过程的随机性降低。我们评估了 PDMS 底物在干细胞增殖和底物介导的分化中的效用。硬度影响多能和分化标记的基因表达，结果表明这些底物系统有可能用于将 hESC 命运引导至早期中胚层谱系。这项研究表明，与可溶性因子相结合，PDMS 底物可能有助于生成特定的分化细胞群。
The pluripotency and self renewing properties of human embryonic stem cells (hESC) make them a valuable tool in the fields of developmental biology, pharmacology and regenerative medicine. Therefore, there exists immense interest in devising strategies for hESC propagation and differentiation. Methods involving simulation of the native stem cell microenvironment, both chemical and physical, have received a lot of attention in recent years. Equally important is evidence that cells can also sense the mechanical properties of their microenvironment. In this study, we test the hypothesis that hESCs accept mechanical cues for differentiation from the substrate by culturing them on flexible polydimethylsiloxane (PDMS) of varying stiffness. PDMS substrates were prepared using available commercial formulations and characterized for stiffness, surface properties and efficiency of cell attachment and proliferation. Across different substrate stiffness, cell numbers, cell attachment and cell surface area were found to be similar. Expression of pluripotency markers decreased with increased time in culture across all PDMS substrates of varying stiffness. Analysis of gene expression of differentiation markers indicates that the differentiation process becomes less stochastic with longer culture times. We evaluated the utility of PDMS substrates for stem cell propagation and substrate mediated differentiation. The stiffness affected gene expression of pluripotent and differentiation markers with results indicating that these substrate systems could potentially be used to direct hESC fate towards early mesodermal lineages. This study suggests that coupled with soluble factors, PDMS substrates could potentially be useful in generating defined populations of differentiated cells.
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影响因子：
3.8
作者：
Li, Lulu;Sharma, Nripen;Langrana, Noshir A.
通讯作者：
Langrana, Noshir A.
影响因子：
5.2
作者：
Gerrard, L;Rodgers, L;Cui, W
通讯作者：
Cui, W
DOI：
10.1126/science.1171643
发表时间：
2009-06-26
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Discher DE;Mooney DJ;Zandstra PW
通讯作者：
Zandstra PW
影响因子：
20.3
作者：
Chadwick, K;Wang, LS;Bhatia, M
通讯作者：
Bhatia, M
影响因子：
4.1
作者：
Hengsberger, S;Kulik, A;Zysset, P
通讯作者：
Zysset, P