Anisotropic stretch-induced hypertrophy in neonatal ventricular myocytes micropatterned on deformable elastomers
Anisotropic stretch-induced hypertrophy in neonatal ventricular myocytes micropatterned on deformable elastomers
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DOI：
10.1002/bit.10506
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发表时间：
2003-03-05
影响因子：
3.8
通讯作者：
McCulloch, AD
中科院分区：
文献类型：
--
作者：
Gopalan, SM;Flaim, C;McCulloch, AD
由于细胞的形状和排列，细胞-基质粘附和细胞-细胞接触都可以影响生长，并且由于体内的机械应变是多轴和各向异性的，我们开发了一种体外系统，用于工程对齐，杆状，新生心肌细胞培养。使用光刻和微流控技术在可变形硅橡胶上以平行线对细胞外基质进行微图案化。通过改变微图案的线宽和胶原蛋白密度来产生汇合的、伸长的、对齐的肌细胞。椭圆形细胞拉伸器将2：1的各向异性应变静态地施加到弹性基底，其中最大拉伸的轴（10%）平行于或横向于肌原纤维。24小时后，与未拉伸对照组相比，与肌细胞平行的主应变没有显著改变肌原纤维积累或心房利钠因子（ANF），连接蛋白-43（Cx-43）或N-钙粘蛋白（通过间接免疫荧光抗体标记和免疫印迹）的表达。相反，10%横向主应变导致肌动蛋白丝连续染色（罗丹明鬼笔环肽）; ANF、Cx-43和N-cadherin的免疫荧光标记增加;以及通过蛋白质印迹上调蛋白质信号强度通过使用微加工和微流体来控制细胞形状和在弹性基底上的排列，我们发现在调节肌节组织、肥大和细胞间连接方面，横向拉伸比纵向拉伸的效果更大。(C)2003 Wiley Periodicals，Inc.
Because cell shape and alignment, cell-matrix adhesion, and cell-cell contact can all affect growth, and because mechanical strains in vivo are multiaxial and anisotropic, we developed an in vitro system for engineering aligned, rod-shaped, neonatal cardiac myocyte cultures. Photolithographic and microfluidic techniques were used to micropattern extracellular matrices in parallel lines on deformable silicone elastomers. Confluent, elongated, aligned myocytes were produced by varying the micropattern line width and collagen density. An elliptical cell stretcher applied 2:1 anisotropic strain statically to the elastic substrate, with the axis of greatest stretch (10%) either parallel or transverse to the myofibrils. After 24 h, the principal strain parallel to myocytes did not significantly alter myofibril accumulation or expression of atrial natriuretic factor (ANF), connexin-43 (Cx-43), or N-cadherin (by indirect immunofluorescent antibody labeling and immunoblotting) compared with unstretched controls. In contrast, 10% transverse principal strain resulted in continuous staining of actin filaments (rhodamine phalloidin); increased immunofluorescent labeling of ANF, Cx-43, and N-cadherin; and up-regulation of protein signal intensity by western blotting By using microfabrication and microfluidics to control cell shape and alignment on an elastic substrate, we found greater effects for transverse than for longitudinal stretch in regulating sarcomere organization, hypertrophy, and cell-to-cell junctions. (C) 2003 Wiley Periodicals, Inc.