Complex interactions between human myoblasts and the surrounding 3D fibrin-based matrix.
Complex interactions between human myoblasts and the surrounding 3D fibrin-based matrix.
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DOI：
10.1371/journal.pone.0036173
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Coirault C
中科院分区：
文献类型：
--
作者：
Chiron S;Tomczak C;Duperray A;Lainé J;Bonne G;Eder A;Hansen A;Eschenhagen T;Verdier C;Coirault C
肌肉细胞与细胞外基质的锚定对于一系列基本生物学过程（包括迁移、生存和分化）至关重要。已经提出三维（3D）培养以提供比常规2D培养更生理的肌肉生长和分化的体外模型。然而，肌细胞粘附和细胞-基质的相互作用的工程肌肉组织仍有待确定。我们已经表征了3D肌肉培养中的细胞-基质相互作用，并分析了它们对细胞分化的影响。将人成肌细胞包埋在两个桩之间的纤维蛋白基质中，培养至汇合，然后诱导分化。成肌细胞在3D中沿凝胶的纵轴沿着。他们显示肌动蛋白应力纤维均匀分布在细胞核周围和薄肌动蛋白丝的皮质网。3D培养中的粘附位点在尺寸上小于刚性2D培养中的粘附位点，但粘附位点蛋白（包括α5整合素和黏着斑蛋白）的表达在3D中高于2D（p<0.05）。成肌细胞和肌管在3D中显示较厚的椭圆形核，而不是2D中的薄盘状核（p<0.001）。在3D中分化动力学更快，如α-辅肌动蛋白和肌球蛋白的较高mRNA浓度所证明的。更重要的是，工程化肌肉组织的弹性模量在增殖过程中从3.5±0.8显著增加到7.4± 4.7kPa（p<0.05），在分化过程中达到12.2± 6.0kPa（p<0.05），从而证明在肌细胞增殖和分化过程中基质刚度增加。总之，我们报告的调制的粘附复合物，肌动蛋白细胞骨架和核形状的3D相比，常规的2D肌肉培养。这些发现指出了肌肉细胞和周围基质之间复杂的相互作用，以及细胞-基质刚度的动态调节。
Anchorage of muscle cells to the extracellular matrix is crucial for a range of fundamental biological processes including migration, survival and differentiation. Three-dimensional (3D) culture has been proposed to provide a more physiological in vitro model of muscle growth and differentiation than routine 2D cultures. However, muscle cell adhesion and cell-matrix interplay of engineered muscle tissue remain to be determined. We have characterized cell-matrix interactions in 3D muscle culture and analyzed their consequences on cell differentiation. Human myoblasts were embedded in a fibrin matrix cast between two posts, cultured until confluence, and then induced to differentiate. Myoblasts in 3D aligned along the longitudinal axis of the gel. They displayed actin stress fibers evenly distributed around the nucleus and a cortical mesh of thin actin filaments. Adhesion sites in 3D were smaller in size than in rigid 2D culture but expression of adhesion site proteins, including α5 integrin and vinculin, was higher in 3D compared with 2D (p<0.05). Myoblasts and myotubes in 3D exhibited thicker and ellipsoid nuclei instead of the thin disk-like shape of the nuclei in 2D (p<0.001). Differentiation kinetics were faster in 3D as demonstrated by higher mRNA concentrations of α-actinin and myosin. More important, the elastic modulus of engineered muscle tissues increased significantly from 3.5±0.8 to 7.4±4.7 kPa during proliferation (p<0.05) and reached 12.2±6.0 kPa during differentiation (p<0.05), thus attesting the increase of matrix stiffness during proliferation and differentiation of the myocytes. In conclusion, we reported modulations of the adhesion complexes, the actin cytoskeleton and nuclear shape in 3D compared with routine 2D muscle culture. These findings point to complex interactions between muscle cells and the surrounding matrix with dynamic regulation of the cell-matrix stiffness.
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影响因子：
64.5
作者：
Choquet, D;Felsenfeld, DP;Sheetz, MP
通讯作者：
Sheetz, MP
影响因子：
3.7
作者：
Matsumoto T;Sasaki J;Alsberg E;Egusa H;Yatani H;Sohmura T
通讯作者：
Sohmura T
影响因子：
4.1
作者：
Hakkinen, Kirsi M.;Harunaga, Jill S.;Yamada, Kenneth M.
通讯作者：
Yamada, Kenneth M.
DOI：
10.1083/jcb.200405004
发表时间：
2004-09-13
期刊：
The Journal of cell biology
影响因子：
--
作者：
Engler AJ;Griffin MA;Sen S;Bönnemann CG;Sweeney HL;Discher DE
通讯作者：
Discher DE
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