Substrate Stiffness Affects Human Keratinocyte Colony Formation.
Substrate Stiffness Affects Human Keratinocyte Colony Formation.
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DOI：
10.1007/s12195-015-0377-8
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发表时间：
2015-03-01
影响因子：
2.8
通讯作者：
Sander, Edward A.
中科院分区：
文献类型：
--
作者：
Zarkoob, Hoda;Bodduluri, Sandeep;Ponnaluri, Sailahari V.;Selby, John C.;Sander, Edward A.
表皮组织和功能对各种病理生理损伤的恢复很大程度上取决于伤口愈合过程中角质形成细胞迁移、增殖和分层的协调。这些过程是由细胞-细胞（桥粒、粘附连接）和细胞-基质（粘着斑、半桥粒）连接以及它们相互连接的细胞骨架丝网络的重新配置介导的。在这项研究中，我们研究了在角质形成细胞培养的钙开关模型触发的新生上皮片形成过程中，基质弹性（硬度）对体外角质形成细胞集落形成的作用。与在硬质（标称 E = 24 kPa）聚丙烯酰胺凝胶上培养的角质形成细胞相比，在胃蛋白酶消化的 I 型胶原包被的柔软（标称 E = 1.2 kPa）聚丙烯酰胺凝胶上培养的角质形成细胞表现出（i）更小的扩散接触面积，（ii）增加的迁移速度，以及（iii）增加的集落形成率，每个集落的细胞更多。通过跟踪嵌入的微球位移进行评估，在软基质上培养的角质形成细胞产生了较大的局部基质变形，这似乎招募了相邻的角质形成细胞加入不断进化的集落。结合观察到的角质形成细胞运动学和基质变形的差异，我们开发了两项临时分析，称为距离等级（DR）和协同半径（RC），这有助于客观地归因于我们所认为的在集落形成过程中在软凝胶和硬凝胶上培养的角质形成细胞日益增加的合作行为。我们假设，在我们的实验中观察到的角质形成细胞集落形成的差异可能是由于局部基质变形产生的细胞-细胞机械信号传导所致，这似乎与位于进化细胞集落外围的角质形成细胞内β4整合素表达的增加相关。
Restoration of epidermal organization and function in response to a variety of pathophysiological insults is critically dependent on coordinated keratinocyte migration, proliferation, and stratification during the process of wound healing. These processes are mediated by the reconfiguration of both cell-cell (desmosomes, adherens junctions) and cell-matrix (focal adhesions, hemidesmosomes) junctions and the cytoskeletal filament networks that they serve to interconnect. In this study, we investigated the role of substrate elasticity (stiffness) on keratinocyte colony formation in vitro during the process of nascent epithelial sheet formation as triggered by the calcium switch model of keratinocyte culture. Keratinocytes cultured on pepsin digested type I collagen coated soft (nominal E = 1.2 kPa) polyacrylamide gels embedded with fluorescent microspheres exhibited (i) smaller spread contact areas, (ii) increased migration velocities, and (iii) increased rates of colony formation with more cells per colony than did keratinocytes cultured on stiff (nominal E = 24 kPa) polyacrylamide gels. As assessed by tracking of embedded microsphere displacements, keratinocytes cultured on soft substrates generated large local substrate deformations that appeared to recruit adjacent keratinocytes into joining an evolving colony. Together with the observed differences in keratinocyte kinematics and substrate deformations, we developed two ad hoc analyses, termed distance rank (DR) and radius of cooperativity (RC), that help to objectively ascribe what we perceive as increasingly cooperative behavior of keratinocytes cultured on soft versus stiff gels during the process of colony formation. We hypothesize that the differences in keratinocyte colony formation observed in our experiments could be due to cell-cell mechanical signaling generated via local substrate deformations that appear to be correlated with the increased expression of β4 integrin within keratinocytes positioned along the periphery of an evolving cell colony.
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影响因子：
41.2
作者：
Kim, Jae Hun;Serra-Picamal, Xavier;Tambe, Dhananjay T.;Zhou, Enhua H.;Park, Chan Young;Sadati, Monirosadat;Park, Jin-Ah;Krishnan, Ramaswamy;Gweon, Bomi;Millet, Emil;Butler, James P.;Trepat, Xavier;Fredberg, Jeffrey J.
通讯作者：
Fredberg, Jeffrey J.
影响因子：
6.5
作者：
Achterberg, Volker F.;Buscemi, Lara;Hinz, Boris
通讯作者：
Hinz, Boris
影响因子：
3.6
作者：
Lange, T S;Kirchberg, J;Scharffetter-Kochanek, K
通讯作者：
Scharffetter-Kochanek, K
DOI：
10.1016/j.jmbbm.2011.08.016
发表时间：
2012-01-01
影响因子：
3.9
作者：
Annaidh, Aisling Ni;Bruyere, Karine;Ottenio, Melanie
通讯作者：
Ottenio, Melanie
DOI：
10.1083/jcb.200810041
发表时间：
2009-02-23
期刊：
The Journal of cell biology
影响因子：
--
作者：
Doyle AD;Wang FW;Matsumoto K;Yamada KM
通讯作者：
Yamada KM