Cell-cell and cell-extracellular matrix interactions regulate embryonic stem cell differentiation
Cell-cell and cell-extracellular matrix interactions regulate embryonic stem cell differentiation
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DOI：
10.1634/stemcells.2006-0419
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发表时间：
2007-03-01
期刊：
影响因子：
5.2
通讯作者：
Margolis, Leonid
中科院分区：
文献类型：
--
作者：
Chen, Silvia S.;Fitzgerald, Wendy;Margolis, Leonid
细胞与细胞外基质（ECM）的相互作用在其生理中起着至关重要的作用。在这里，我们试图确定外源性和内源性ECM在非人灵长类ESCs分化中的作用。我们利用实时聚合酶链反应和免疫组织化学技术从谱系基因mRNA和蛋白质的表达来评估细胞分化。我们发现附着在黏附性强的胶原上并扩散的ESCs不能有效地分化，而在黏附性较差的基质上，ESCs形成聚集体并沿中胚层尤其是内胚层分化。血统。为了进一步减少ESC与底物的附着，我们将它们分别培养在不粘附的琼脂糖上或悬浮中。在这两种情况下，ESCs形成聚集体并沿内胚层和中胚层谱系有效分化，最显著的是分化为心肌细胞。由这种分化的ESCs形成的聚集体开始以每分钟50-100次的频率跳动，并持续跳动约一个月。尽管存在外源性ECM，但ESCs的存活和分化仍依赖于内源性ECM，因为内源性胶原的抑制会导致ESCs的逐渐丧失，而简单的基质（如I型胶原）和复杂的基质Matrigel都无法挽救这些细胞。总之，对各种ECM和非生物底物的粘附性决定了ESCs的分化，这样有效的细胞-细胞聚集，加上效率较低的细胞附着和扩散，导致了更有效的细胞分化。
Cell interactions with the extracellular matrix (ECM) play a critical role in their physiology. Here, we sought to determine the role of exogenous and endogenous ECM in the differentiation of nonhuman primate ESCs. We evaluated cell differentiation from expression of lineage gene mRNA and proteins using real-time polymerase chain reaction and immunohistochemistry. We found that ESCs that attached to and spread upon highly adhesive collagen do not differentiate efficiently, whereas on the less adhesive Matrigel, ESCs form aggregates and differentiate along mesoderm and especially endoderm. lineages. To further decrease ESC attachment to the substrate, we cultured them either on nonadhesive agarose or in suspension. In both cases, ESCs formed aggregates and efficiently differentiated along endoderm and mesoderm lineages, most strikingly into cardiomyocytes. Aggregates formed by thus-differentiated ESCs started to beat with a frequency of 50-100 beats per minute and continued to beat for approximately a month. In spite of the presence of exogenous ECM, ESCs were dependent on endogenous ECM for their survival and differentiation, as the inhibition of endogenous collagen induced a gradual loss of ESCs and neither a simple matrix, such as type I collagen, nor the complex matrix Matrigel was able to rescue these cells. In conclusion, adhesiveness to various ECM and nonbiological substrates determines the differentiation of ESCs in such a way that efficient cell-cell aggregation, together with less efficient cell attachment and spreading, results in more efficient cell differentiation.