Soft substrates promote homogeneous self-renewal of embryonic stem cells via downregulating cell-matrix tractions.
Soft substrates promote homogeneous self-renewal of embryonic stem cells via downregulating cell-matrix tractions.
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DOI：
10.1371/journal.pone.0015655
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发表时间：
2010-12-13
期刊：
影响因子：
3.7
通讯作者：
Tanaka TS
中科院分区：
文献类型：
--
作者：
Chowdhury F;Li Y;Poh YC;Yokohama-Tamaki T;Wang N;Tanaka TS
维持未分化的小鼠胚胎干细胞（mESC）培养一直是一个主要挑战，因为在白血病抑制因子（LIF）条件下培养的mESC表现出自发分化、多能性基因和特化细胞基因的波动表达。在这里，我们表明，与接种在常规刚性基底上的mESC形成鲜明对比的是，在与mESC的固有刚度相匹配的软基底上培养的mESC，并且在不存在外源LIF的情况下培养5天，令人惊讶地仍然产生均匀的未分化集落，保持高水平的Oct 3/4、Nanog和碱性磷酸酶（AP）活性，并且有效地形成胚状体和畸胎瘤。在不含外源LIF的软基质上培养的不同的mESC系保持了产生具有相对高水平的Oct 3/4和AP活性的同质未分化集落的能力，最高达至少15代，表明这种软基质方法适用于不同mESC系的长期培养。在这些没有LIF的软基质上的mESC集落产生低细胞-基质牵引力和低刚度。牵引力和刚度的殖民地增加与基板刚度，伴随着下调Oct 3/4的表达。我们的研究结果表明，mESC自我更新和多能性可以保持均匀的软基板上通过生物物理机制，促进低细胞基质牵引的产生。
Maintaining undifferentiated mouse embryonic stem cell (mESC) culture has been a major challenge as mESCs cultured in Leukemia Inhibitory Factor (LIF) conditions exhibit spontaneous differentiation, fluctuating expression of pluripotency genes, and genes of specialized cells. Here we show that, in sharp contrast to the mESCs seeded on the conventional rigid substrates, the mESCs cultured on the soft substrates that match the intrinsic stiffness of the mESCs and in the absence of exogenous LIF for 5 days, surprisingly still generated homogeneous undifferentiated colonies, maintained high levels of Oct3/4, Nanog, and Alkaline Phosphatase (AP) activities, and formed embryoid bodies and teratomas efficiently. A different line of mESCs, cultured on the soft substrates without exogenous LIF, maintained the capacity of generating homogeneous undifferentiated colonies with relatively high levels of Oct3/4 and AP activities, up to at least 15 passages, suggesting that this soft substrate approach applies to long term culture of different mESC lines. mESC colonies on these soft substrates without LIF generated low cell-matrix tractions and low stiffness. Both tractions and stiffness of the colonies increased with substrate stiffness, accompanied by downregulation of Oct3/4 expression. Our findings demonstrate that mESC self-renewal and pluripotency can be maintained homogeneously on soft substrates via the biophysical mechanism of facilitating generation of low cell-matrix tractions.
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DOI：
10.1083/jcb.201001149
发表时间：
2010-06-28
期刊：
The Journal of cell biology
影响因子：
--
作者：
le Duc Q;Shi Q;Blonk I;Sonnenberg A;Wang N;Leckband D;de Rooij J
通讯作者：
de Rooij J
DOI：
10.1083/jcb.201006094
发表时间：
2010-11-01
期刊：
The Journal of cell biology
影响因子：
--
作者：
Li D;Zhou J;Wang L;Shin ME;Su P;Lei X;Kuang H;Guo W;Yang H;Cheng L;Tanaka TS;Leckband DE;Reynolds AB;Duan E;Wang F
通讯作者：
Wang F
影响因子：
23.9
作者：
Hayashi K;de Sousa Lopes SMC;Tang F;Lao K;Surani MA
通讯作者：
Surani MA
影响因子：
4.8
作者：
Duval, D;Reinhardt, S;Boeuf, H
通讯作者：
Boeuf, H
影响因子：
3.4
作者：
Engler, A;Bacakova, L;Discher, D
通讯作者：
Discher, D